def splitRegions(inputGFF,tssCollection):
#if even a single coordinate is shared with the +/-1kb
splitGFF = []
for line in inputGFF:
chrom = line[0]
regionID = line[1]
lineLocus = utils.Locus(line[0],line[3],line[4],'.')
overlappingLoci = tssCollection.getOverlap(lineLocus)
if len(overlappingLoci) > 0: #case where a tss Overlap
#identify the parts of the line locus that are contained
localTSSCollection = utils.LocusCollection(overlappingLoci,50)
overlappingCoords = lineLocus.coords()
for tssLocus in overlappingLoci:
overlappingCoords += tssLocus.coords()
overlappingCoords = utils.uniquify(overlappingCoords)
overlappingCoords.sort()
#you need to hack and slash add 1 to the last coordinate of the overlappingCoords
overlappingCoords[-1] +=1
i = 0
regionTicker = 1
while i < (len(overlappingCoords)-1):
start = int(overlappingCoords[i])
stop = int(overlappingCoords[(i+1)])-1
if (stop - start) < 50: #this eliminates really tiny regions
i+=1
continue
splitLocus = utils.Locus(chrom,start+1,stop,'.')
if lineLocus.overlaps(splitLocus): #has to be a mycn site
newID = '%s_%s' % (regionID,regionTicker)
tssStatus = 0
if localTSSCollection.getOverlap(splitLocus):
tssStatus = 1
splitGFFLine = [chrom,newID,newID,start,stop,'','.',tssStatus,newID]
splitGFF.append(splitGFFLine)
regionTicker+=1
i+=1
else:
line[7] = 0
splitGFF.append(line)
return splitGFF
def makeBedCollection(bedFileList):
'''
takes in a list of bedFiles and makes a single huge collection
each locus has as its ID the name of the bed file
'''
bedLoci = []
print("MAKING BED COLLECTION FOR:")
for bedFile in bedFileList:
bedName = bedFile.split('/')[-1].split('.')[0]
print(bedName)
bed = utils.parseTable(bedFile, '\t')
for line in bed:
if len(line) >= 3:
#check that line[0]
if line[0][0:3] == 'chr':
try:
coords = [int(line[1]), int(line[2])]
bedLocus = utils.Locus(line[0], min(coords),
max(coords), '.', bedName)
bedLoci.append(bedLocus)
except ValueError:
pass
print("IDENTIFIED %s BED REGIONS" % (len(bedLoci)))
return utils.LocusCollection(bedLoci, 50)
def findValleys(TFtoEnhancerDict, bamFile, projectName, projectFolder, cutoff = 0.2):
'''
takes in the super dict
returns a dictionary of refseqs with all valley loci that are associated
'''
print 'IDENTIFYING VALLEYS IN SUPER ENHANCERS'
valleyBED = []
valleyDict = {}
for gene in TFtoEnhancerDict.keys():
valleyDict[gene] = []
print gene
for region in TFtoEnhancerDict[gene]:
scoreArray = scoreValley(region, bamFile, projectName, projectFolder)
for index,score in enumerate(scoreArray):
if score > cutoff:
valley = utils.Locus(region.chr(), region.start() + index*10,
region.start() + (index+1)*10, '.')
valleyDict[gene].append(valley)
stitchedValleys = stitchValleys(valleyDict[gene])
for valley in stitchedValleys:
valleyBED.append([valley.chr(), valley.start(), valley.end()])
valleyDict[gene] = stitchedValleys
bedfilename = projectFolder + projectName + '_valleys.bed'
utils.unParseTable(valleyBED, bedfilename, '\t')
print bedfilename
return bedfilename
def main():
projectFolder = '/storage/goodell/projects/jmreyes/amish_ayala/'
bedFolder = projectFolder+'bed/'
wtCanyonBed = bedFolder+'canyon_WT_sizeSelected.bed'
mutCanyonBed = bedFolder+'canyon_Mut_sizeSelected.bed'
wtCanyonLocusCollection = utils.LocusCollection([utils.Locus(x[0], x[1], x[2], '.', 'wt_'+str(x[0])+':'+str(x[1])+'-'+str(x[2])) for x in utils.parseTable(wtCanyonBed, '\t')])
mutCanyonLocusCollection = utils.LocusCollection([utils.Locus(x[0], x[1], x[2], '.', 'mut_'+str(x[0])+':'+str(x[1])+'-'+str(x[2])) for x in utils.parseTable(mutCanyonBed, '\t')])
overlappingCanyons = []
wtExpansion = []
mutExpansion = []
wtUnique = []
mutUnique = []
overlapCounter = 0
mutOverlap = 0
for locus in wtCanyonLocusCollection.getLoci():
wtMutOverlap = mutCanyonLocusCollection.getOverlap(locus, 'both')
if len(wtMutOverlap) > 0:
overlapCounter += 1
for overlap in wtMutOverlap:
newLine = [locus.chr(), locus.start(), locus.end(), locus.end()-locus.start(), overlap.chr(), overlap.start(), overlap.end(), overlap.end()-overlap.start()]
wtLength = locus.end()-locus.start()
mutLength = overlap.end()-overlap.start()
if mutLength > wtLength:
mutExpansion.append(newLine)
elif wtLength > mutLength:
wtExpansion.append(newLine)
else:
wtUnique.append(locus)
for locus in mutCanyonLocusCollection.getLoci():
mutWTOverlap = wtCanyonLocusCollection.getOverlap(locus, 'both')
if len(mutWTOverlap) > 0:
mutOverlap += 1
else:
mutUnique.append(locus)
print len(mutExpansion)
print len(wtExpansion)
utils.unParseTable(mutExpansion, projectFolder+'tables/MUT_canyons_expanded.txt', '\t')
utils.unParseTable(wtExpansion, projectFolder+'tables/WT_canyons_expanded.txt', '\t')
def assignEnhancerRank(enhancerToGeneFile,
enhancerFile1,
enhancerFile2,
name1,
name2,
rankOutput=''):
'''
for all genes in the enhancerToGene Table, assigns the highest overlapping ranked enhancer in the other tables
'''
enhancerToGene = utils.parseTable(enhancerToGeneFile, '\t')
enhancerCollection1 = makeSECollection(enhancerFile1, name1, False)
enhancerCollection2 = makeSECollection(enhancerFile2, name2, False)
enhancerDict1 = makeSEDict(enhancerFile1, name1, False)
enhancerDict2 = makeSEDict(enhancerFile2, name2, False)
#we're going to update the enhancerToGeneTable
enhancerToGene[0] += ['%s_rank' % name1, '%s_rank' % name2]
for i in range(1, len(enhancerToGene)):
line = enhancerToGene[i]
locusLine = utils.Locus(line[1], line[2], line[3], '.', line[0])
#if the enhancer doesn't exist, its ranking is dead last on the enhancer list
enhancer1Overlap = enhancerCollection1.getOverlap(locusLine, 'both')
if len(enhancer1Overlap) == 0:
enhancer1Rank = len(enhancerCollection1)
else:
rankList1 = [
enhancerDict1[x.ID()]['rank'] for x in enhancer1Overlap
]
enhancer1Rank = min(rankList1)
enhancer2Overlap = enhancerCollection2.getOverlap(locusLine, 'both')
if len(enhancer2Overlap) == 0:
enhancer2Rank = len(enhancerCollection2)
else:
rankList2 = [
enhancerDict2[x.ID()]['rank'] for x in enhancer2Overlap
]
enhancer2Rank = min(rankList2)
enhancerToGene[i] += [enhancer1Rank, enhancer2Rank]
if len(rankOutput) == 0:
return enhancerToGene
else:
utils.unParseTable(enhancerToGene, rankOutput, '\t')
def generateSubpeakFASTA(TFtoEnhancerDict, subpeaks, genomeDirectory,
projectName, projectFolder, constExtension):
'''
from a BED file of constituents
generate a FASTA for the consituients contained within the canidate supers
'''
subpeakDict = {}
subpeakBED = [['track name=' + projectName + ' color=204,0,204']]
subpeakTable = utils.parseTable(subpeaks, '\t')
subpeakLoci = [
utils.Locus(l[0], int(l[1]), int(l[2]), '.') for l in subpeakTable
]
subpeakCollection = utils.LocusCollection(subpeakLoci, 50)
for gene in TFtoEnhancerDict.keys():
subpeakDict[gene] = []
for region in TFtoEnhancerDict[gene]:
overlaps = subpeakCollection.getOverlap(region)
extendedOverlaps = [
utils.makeSearchLocus(x, constExtension, constExtension)
for x in overlaps
]
overlapCollectionTemp = utils.LocusCollection(extendedOverlaps, 50)
overlapCollection = overlapCollectionTemp.stitchCollection()
for overlap in overlapCollection.getLoci():
subpeakBED.append(
[overlap.chr(),
overlap.start(),
overlap.end()])
subpeakDict[gene].append(overlap)
bedfilename = projectFolder + projectName + '_subpeaks.bed'
utils.unParseTable(subpeakBED, bedfilename, '\t')
fasta = []
for gene in subpeakDict:
for subpeak in subpeakDict[gene]:
fastaTitle = gene + '|' + subpeak.chr() + '|' + str(
subpeak.start()) + '|' + str(subpeak.end())
fastaLine = utils.fetchSeq(genomeDirectory, subpeak.chr(),
int(subpeak.start() + 1),
int(subpeak.end() + 1))
fasta.append('>' + fastaTitle)
fasta.append(upper(fastaLine))
outname = projectFolder + projectName + '_SUBPEAKS.fa'
utils.unParseTable(fasta, outname, '')
def mapGFFLineToBed(gffLine, outFolder, nBins, bedCollection, header=''):
'''
for every line produces a file with all of the rectangles to draw
'''
if len(header) == 0:
gffString = '%s_%s_%s_%s' % (gffLine[0], gffLine[6], gffLine[3],
gffLine[4])
else:
gffString = header
diagramTable = [[0, 0, 0, 0]]
nameTable = [['', 0, 0]]
gffLocus = utils.Locus(gffLine[0], int(gffLine[3]), int(gffLine[4]),
gffLine[6], gffLine[1])
scaleFactor = float(nBins) / gffLocus.len()
# plotting buffer for diagrams
# plotBuffer = int(gffLocus.len() / float(nBins) * 20) # UNUSED (?)
overlapLoci = bedCollection.getOverlap(gffLocus, sense='both')
print("IDENTIFIED %s OVERLAPPING BED LOCI FOR REGION %s" %
(len(overlapLoci), gffLine))
# since beds come from multiple sources, we want to figure out how to offset them
offsetDict = {} # this will store each ID name
bedNamesList = utils.uniquify([locus.ID() for locus in overlapLoci])
bedNamesList.sort()
for i in range(len(bedNamesList)):
offsetDict[bedNamesList[
i]] = 2 * i # offsets different categories of bed regions
if gffLine[6] == '-':
refPoint = int(gffLine[4])
else:
refPoint = int(gffLine[3])
# fill out the name table
for name in bedNamesList:
offset = offsetDict[name]
nameTable.append([name, 0, 0.0 - offset])
for bedLocus in overlapLoci:
offset = offsetDict[bedLocus.ID()]
[start,
stop] = [abs(x - refPoint) * scaleFactor for x in bedLocus.coords()]
diagramTable.append([start, -0.5 - offset, stop, 0.5 - offset])
utils.unParseTable(diagramTable,
outFolder + gffString + '_bedDiagramTemp.txt', '\t')
utils.unParseTable(nameTable, outFolder + gffString + '_bedNameTemp.txt',
'\t')
def findValleys(gene_to_enhancer_dict,
bamFileList,
projectName,
projectFolder,
cutoff=0.2):
'''
takes in the super dict
returns a dictionary of refseqs with all valley loci that are associated
returns 2 kinds of bed files...
1 = all
'''
#first make the bamDict
all_valley_bed = []
valleyDict = {}
#start w/ a bamFileList and make a list of bam type objects
bam_list = [utils.Bam(bam_path) for bam_path in bamFileList]
max_read_length = max([bam.getReadLengths()[0] for bam in bam_list])
gene_list = gene_to_enhancer_dict.keys()
gene_list.sort()
ticker = 0
print('number of regions processed:')
for gene in gene_list:
valleyDict[gene] = []
for region in gene_to_enhancer_dict[gene]:
if ticker % 100 == 0:
print(ticker)
ticker += 1
scoreArray = scoreValley(region, bam_list, max_read_length,
projectName, projectFolder)
for index, score in enumerate(scoreArray):
if score > cutoff:
valley = utils.Locus(region.chr(),
region.start() + index * 10,
region.start() + (index + 1) * 10,
'.')
valleyDict[gene].append(valley)
stitchedValleys = stitchValleys(valleyDict[gene])
for valley in stitchedValleys:
all_valley_bed.append([valley.chr(), valley.start(), valley.end()])
valleyDict[gene] = stitchedValleys
all_bed_path = projectFolder + projectName + '_all_valleys.bed'
utils.unParseTable(all_valley_bed, all_bed_path, '\t')
return all_bed_path
def createSuperLoci(superTable, Enumber='super'):
'''
takes as input a ROSE SuperEnhancer table
output a table of loci for SuperEnhancers
'''
print 'CREATING SUPER-ENHANCER LOCUS COLLECTION'
output = []
if Enumber == 'super':
for line in superTable[6:]:
if line[-1] == '1':
locus = utils.Locus(line[1], line[2], line[3], '.', line[0], (float(line[6])-float(line[7])))
output.append(locus)
else:
end = 6+int(Enumber)
for line in superTable[6:end]:
locus = utils.Locus(line[1], line[2], line[3], '.', line[0], (float(line[6])-float(line[7])))
output.append(locus)
return output
def makePeakGFFs(peak_path_list):
'''
makes a stitched gff for all MYC bound TSS and Distal regions across all datasets
'''
#setting the output
tss_gff_path = '%sHG19_MYC_TSS_REGIONS_-0_+0.gff' % (gffFolder)
distal_gff_path = '%sHG19_MYC_DISTAL_REGIONS_-0_+0.gff' % (gffFolder)
#check to see if already done
if utils.checkOutput(tss_gff_path,0.1,0.1) and utils.checkOutput(distal_gff_path,0.1,0.1):
print('OUTPUT FOUND AT %s and %s' % (tss_gff_path,distal_gff_path))
return tss_gff_path,distal_gff_path
#emtpy loci lists to hold everything
tss_loci = []
distal_loci = []
for peak_path in peak_path_list:
print('processing %s' % (peak_path))
peak_table= utils.parseTable(peak_path,'\t')
for line in peak_table[1:]:
peak_locus = utils.Locus(line[1],line[2],line[3],'.')
if int(line[5]) == 0:
distal_loci.append(peak_locus)
else:
tss_loci.append(peak_locus)
#now combind the loci
print('stitching loci')
distal_collection = utils.LocusCollection(distal_loci,50)
tss_collection = utils.LocusCollection(tss_loci,50)
stitched_distal_collection = distal_collection.stitchCollection()
stitched_tss_collection = tss_collection.stitchCollection()
#now make the gffs
distal_gff= utils.locusCollectionToGFF(distal_collection)
tss_gff= utils.locusCollectionToGFF(tss_collection)
#now write to disk
utils.unParseTable(distal_gff,distal_gff_path,'\t')
utils.unParseTable(tss_gff,tss_gff_path,'\t')
return tss_gff_path,distal_gff_path
def make_mycn_regions(conserved_rank_path):
'''
takes conserved NB MYCN regions
then creates a bed and gff of regions
'''
conserved_rank_table = utils.parseTable(conserved_rank_path,'\t')
mycn_gff = []
mycn_flank_gff = []
mycn_bed = []
mycn_flank_bed = []
for line in conserved_rank_table[1:]:
locus_line = utils.Locus(line[1],line[2],line[3],'.')
if int(line[3]) < int(line[2]):
print('uh oh')
print(line)
gff_line = [line[1],line[0],'',line[2],line[3],'','.','',line[0]]
bed_line = [line[1],line[2],line[3],line[0]]
mycn_gff.append(gff_line)
mycn_bed.append(bed_line)
gff_flank_line = [line[1],line[0],'',int(line[2])-500,int(line[3])+500,'','.','',line[0]]
bed_flank_line = [line[1],int(line[2])-500,int(line[3])+500,line[0]]
mycn_flank_gff.append(gff_flank_line)
mycn_flank_bed.append(bed_flank_line)
mycn_gff_path = '%sHG19_NB_MYCN_CONSERVED_-0_+0.gff' % (gffFolder)
mycn_flank_gff_path = '%sHG19_NB_MYCN_CONSERVED_-500_+500.gff' % (gffFolder)
mycn_bed_path = '%sHG19_NB_MYCN_CONSERVED_-0_+0.bed' % (bedFolder)
mycn_flank_bed_path = '%sHG19_NB_MYCN_CONSERVED_-500_+500.bed' % (bedFolder)
#writing to disk
utils.unParseTable(mycn_gff,mycn_gff_path,'\t')
utils.unParseTable(mycn_flank_gff,mycn_flank_gff_path,'\t')
utils.unParseTable(mycn_bed,mycn_bed_path,'\t')
utils.unParseTable(mycn_flank_bed,mycn_flank_bed_path,'\t')
print(mycn_gff_path)
print(mycn_flank_gff_path)
print(mycn_bed_path)
print(mycn_flank_bed_path)
return mycn_gff_path,mycn_flank_gff_path
def mapBamToGFFLine(bamFile, MMR, name, gffLine, color, nBins, sense='both', extension=200):
'''maps reads from a bam to a gff'''
print('using a MMR/scaling denominator value of %s' % (MMR))
line = gffLine[0:9]
gffLocus = utils.Locus(line[0], int(line[3]), int(line[4]), line[6], line[1])
# setting up the output clusterline
colorLine = color
bamName = bamFile.split('/')[-1]
clusterLine = [bamName, gffLocus.ID(), name, gffLocus.__str__()] + colorLine
binSize = gffLocus.len() / nBins
# some regions will be too short to get info on
# we just kick these back and abandon them
if binSize == 0:
clusterLine += ['NA'] * int(nBins)
return clusterLine
# flippy flip if sense is negative
senseTrans = string.maketrans('-+.', '+-+')
if sense == '-':
bamSense = string.translate(gffLocus.sense(), senseTrans)
elif sense == '+':
bamSense = gffLocus.sense()
else:
bamSense = '.'
# using the bamLiquidator to get the readstring
# print('using nBin of %s' % nBin)
bamCommand = "%s %s %s %s %s %s %s %s" % (bamliquidatorString, bamFile, gffLocus.chr(), gffLocus.start(), gffLocus.end(), bamSense, nBins, extension)
# print(bamCommand)
getReads = subprocess.Popen(bamCommand, stdin=subprocess.PIPE, stderr=subprocess.PIPE, stdout=subprocess.PIPE, shell=True)
readString = getReads.communicate()
denList = readString[0].split('\n')[:-1]
# flip the denList if the actual gff region is -
if gffLocus.sense() == '-':
denList = denList[::-1]
# converting from units of total bp of read sequence per bin to rpm/bp
denList = [round(float(x) / binSize / MMR, 4) for x in denList]
clusterLine += denList
return clusterLine
def generateSubpeakFASTA(TFandSuperDict, subpeaks, genomeDirectory, projectName, projectFolder, motifExtension):
'''
takes as input a BED file of constituents
outputs a FASTA file of merged extended super-enhancer consituents and associated formated name
'''
print 'MAKE FASTA'
subpeakDict = {}
subpeakBED = [['track name=' + projectName + ' color=204,0,204']]
subpeakTable = utils.parseTable(subpeaks, '\t')
subpeakLoci = [utils.Locus(l[0], int(l[1]), int(l[2]), '.') for l in subpeakTable]
subpeakCollection = utils.LocusCollection(subpeakLoci, 50)
for gene in TFandSuperDict.keys():
subpeakDict[gene] = []
for region in TFandSuperDict[gene]:
overlaps = subpeakCollection.getOverlap(region)
extendedOverlaps = [utils.makeSearchLocus(x, motifExtension, motifExtension) for x in overlaps]
overlapCollectionTemp = utils.LocusCollection(extendedOverlaps, 50)
overlapCollection = overlapCollectionTemp.stitchCollection()
for overlap in overlapCollection.getLoci():
subpeakBED.append([overlap.chr(), overlap.start(), overlap.end()])
subpeakDict[gene].append(overlap)
bedfilename = projectFolder + projectName + '_subpeaks.bed'
utils.unParseTable(subpeakBED, bedfilename, '\t')
fasta = []
for gene in subpeakDict:
for subpeak in subpeakDict[gene]:
fastaTitle = gene + '|' + subpeak.chr() + '|' + str(subpeak.start()) + '|' + str(subpeak.end())
fastaLine = utils.fetchSeq(genomeDirectory, subpeak.chr(), int(subpeak.start()+1), int(subpeak.end()+1))
fasta.append('>' + fastaTitle)
fasta.append(upper(fastaLine))
# Output the fasta file of extended SE constituents
outname = projectFolder + projectName + '_SUBPEAKS.fa'
utils.unParseTable(fasta, outname, '')
def makeSECollection(enhancerFile, name, superOnly=True):
'''
returns a locus collection from a super table
top gives the number of rows
'''
enhancerTable = utils.parseTable(enhancerFile, '\t')
enhancerLoci = []
for line in enhancerTable:
if line[0][0] == '#' or line[0][0] == 'R':
continue
else:
if superOnly and int(line[-1]) == 0:
break
enhancerLoci.append(
utils.Locus(line[1], line[2], line[3], '.',
name + '_' + line[0]))
return utils.LocusCollection(enhancerLoci, 50)
def makeSECollection(enhancerFile,name,top=0):
'''
returns a locus collection from a super table
top gives the number of rows
'''
enhancerTable = utils.parseTable(enhancerFile,'\t')
superLoci = []
ticker = 0
for line in enhancerTable:
if line[0][0] == '#' or line[0][0] == 'R':
continue
else:
ticker+=1
superLoci.append(utils.Locus(line[1],line[2],line[3],'.',name+'_'+line[0]))
if ticker == top:
break
return utils.LocusCollection(superLoci,50)
def mapCollection(stitchedCollection, referenceCollection, bamFileList,
mappedFolder, output, refName):
'''
makes a table of factor density in a stitched locus and ranks table by number of loci stitched together
'''
print('FORMATTING TABLE')
loci = stitchedCollection.getLoci()
locusTable = [[
'REGION_ID', 'CHROM', 'START', 'STOP', 'NUM_LOCI', 'CONSTITUENT_SIZE'
]]
lociLenList = []
# strip out any that are in chrY
for locus in list(loci):
if locus.chr() == 'chrY':
loci.remove(locus)
for locus in loci:
# numLociList.append(int(stitchLocus.ID().split('_')[1]))
lociLenList.append(locus.len())
# numOrder = order(numLociList,decreasing=True)
lenOrder = utils.order(lociLenList, decreasing=True)
ticker = 0
for i in lenOrder:
ticker += 1
if ticker % 1000 == 0:
print(ticker)
locus = loci[i]
# First get the size of the enriched regions within the stitched locus
refEnrichSize = 0
refOverlappingLoci = referenceCollection.getOverlap(locus, 'both')
for refLocus in refOverlappingLoci:
refEnrichSize += refLocus.len()
try:
stitchCount = int(locus.ID().split('_')[0])
except ValueError:
stitchCount = 1
coords = [int(x) for x in locus.coords()]
locusTable.append([
locus.ID(),
locus.chr(),
min(coords),
max(coords), stitchCount, refEnrichSize
])
print('GETTING MAPPED DATA')
print("USING A BAMFILE LIST:")
print(bamFileList)
for bamFile in bamFileList:
bamFileName = bamFile.split('/')[-1]
print('GETTING MAPPING DATA FOR %s' % bamFile)
# assumes standard convention for naming enriched region gffs
# opening up the mapped GFF
print('OPENING %s%s_%s_MAPPED/matrix.txt' %
(mappedFolder, refName, bamFileName))
mappedGFF = utils.parseTable(
'%s%s_%s_MAPPED/matrix.txt' % (mappedFolder, refName, bamFileName),
'\t')
signalDict = defaultdict(float)
print('MAKING SIGNAL DICT FOR %s' % (bamFile))
mappedLoci = []
for line in mappedGFF[1:]:
chrom = line[1].split('(')[0]
start = int(line[1].split(':')[-1].split('-')[0])
end = int(line[1].split(':')[-1].split('-')[1])
mappedLoci.append(utils.Locus(chrom, start, end, '.', line[0]))
try:
signalDict[line[0]] = float(line[2]) * (abs(end - start))
except ValueError:
print('WARNING NO SIGNAL FOR LINE:')
print(line)
continue
mappedCollection = utils.LocusCollection(mappedLoci, 500)
locusTable[0].append(bamFileName)
for i in range(1, len(locusTable)):
signal = 0.0
line = locusTable[i]
lineLocus = utils.Locus(line[1], line[2], line[3], '.')
overlappingRegions = mappedCollection.getOverlap(lineLocus,
sense='both')
for region in overlappingRegions:
signal += signalDict[region.ID()]
locusTable[i].append(signal)
utils.unParseTable(locusTable, output, '\t')
def mapEnhancerToGene(annotFile,enhancerFile,transcribedFile='',uniqueGenes=True,searchWindow =50000,noFormatTable = False):
'''
maps genes to enhancers. if uniqueGenes, reduces to gene name only. Otherwise, gives for each refseq
'''
startDict = utils.makeStartDict(annotFile)
enhancerTable = utils.parseTable(enhancerFile,'\t')
#internal parameter for debugging
byRefseq = False
if len(transcribedFile) > 0:
transcribedTable = utils.parseTable(transcribedFile,'\t')
transcribedGenes = [line[1] for line in transcribedTable]
else:
transcribedGenes = startDict.keys()
print('MAKING TRANSCRIPT COLLECTION')
transcribedCollection = utils.makeTranscriptCollection(annotFile,0,0,500,transcribedGenes)
print('MAKING TSS COLLECTION')
tssLoci = []
for geneID in transcribedGenes:
tssLoci.append(utils.makeTSSLocus(geneID,startDict,0,0))
#this turns the tssLoci list into a LocusCollection
#50 is the internal parameter for LocusCollection and doesn't really matter
tssCollection = utils.LocusCollection(tssLoci,50)
geneDict = {'overlapping':defaultdict(list),'proximal':defaultdict(list)}
#dictionaries to hold ranks and superstatus of gene nearby enhancers
rankDict = defaultdict(list)
superDict= defaultdict(list)
#list of all genes that appear in this analysis
overallGeneList = []
if noFormatTable:
#set up the output tables
#first by enhancer
enhancerToGeneTable = [enhancerTable[0]+['OVERLAP_GENES','PROXIMAL_GENES','CLOSEST_GENE']]
else:
#set up the output tables
#first by enhancer
enhancerToGeneTable = [enhancerTable[0][0:9]+['OVERLAP_GENES','PROXIMAL_GENES','CLOSEST_GENE'] + enhancerTable[5][-2:]]
#next by gene
geneToEnhancerTable = [['GENE_NAME','REFSEQ_ID','PROXIMAL_ENHANCERS']]
#next make the gene to enhancer table
geneToEnhancerTable = [['GENE_NAME','REFSEQ_ID','PROXIMAL_ENHANCERS','ENHANCER_RANKS','IS_SUPER']]
for line in enhancerTable:
if line[0][0] =='#' or line[0][0] == 'R':
continue
enhancerString = '%s:%s-%s' % (line[1],line[2],line[3])
enhancerLocus = utils.Locus(line[1],line[2],line[3],'.',line[0])
#overlapping genes are transcribed genes whose transcript is directly in the stitchedLocus
overlappingLoci = transcribedCollection.getOverlap(enhancerLocus,'both')
overlappingGenes =[]
for overlapLocus in overlappingLoci:
overlappingGenes.append(overlapLocus.ID())
#proximalGenes are transcribed genes where the tss is within 50kb of the boundary of the stitched loci
proximalLoci = tssCollection.getOverlap(utils.makeSearchLocus(enhancerLocus,searchWindow,searchWindow),'both')
proximalGenes =[]
for proxLocus in proximalLoci:
proximalGenes.append(proxLocus.ID())
distalLoci = tssCollection.getOverlap(utils.makeSearchLocus(enhancerLocus,1000000,1000000),'both')
distalGenes =[]
for proxLocus in distalLoci:
distalGenes.append(proxLocus.ID())
overlappingGenes = utils.uniquify(overlappingGenes)
proximalGenes = utils.uniquify(proximalGenes)
distalGenes = utils.uniquify(distalGenes)
allEnhancerGenes = overlappingGenes + proximalGenes + distalGenes
#these checks make sure each gene list is unique.
#technically it is possible for a gene to be overlapping, but not proximal since the
#gene could be longer than the 50kb window, but we'll let that slide here
for refID in overlappingGenes:
if proximalGenes.count(refID) == 1:
proximalGenes.remove(refID)
for refID in proximalGenes:
if distalGenes.count(refID) == 1:
distalGenes.remove(refID)
#Now find the closest gene
if len(allEnhancerGenes) == 0:
closestGene = ''
else:
#get enhancerCenter
enhancerCenter = (int(line[2]) + int(line[3]))/2
#get absolute distance to enhancer center
distList = [abs(enhancerCenter - startDict[geneID]['start'][0]) for geneID in allEnhancerGenes]
#get the ID and convert to name
closestGene = startDict[allEnhancerGenes[distList.index(min(distList))]]['name']
#NOW WRITE THE ROW FOR THE ENHANCER TABLE
if noFormatTable:
newEnhancerLine = list(line)
newEnhancerLine.append(join(utils.uniquify([startDict[x]['name'] for x in overlappingGenes]),','))
newEnhancerLine.append(join(utils.uniquify([startDict[x]['name'] for x in proximalGenes]),','))
newEnhancerLine.append(closestGene)
else:
newEnhancerLine = line[0:9]
newEnhancerLine.append(join(utils.uniquify([startDict[x]['name'] for x in overlappingGenes]),','))
newEnhancerLine.append(join(utils.uniquify([startDict[x]['name'] for x in proximalGenes]),','))
newEnhancerLine.append(closestGene)
newEnhancerLine += line[-2:]
enhancerToGeneTable.append(newEnhancerLine)
#Now grab all overlapping and proximal genes for the gene ordered table
overallGeneList +=overlappingGenes
for refID in overlappingGenes:
geneDict['overlapping'][refID].append(enhancerString)
rankDict[refID].append(int(line[-2]))
superDict[refID].append(int(line[-1]))
overallGeneList+=proximalGenes
for refID in proximalGenes:
geneDict['proximal'][refID].append(enhancerString)
rankDict[refID].append(int(line[-2]))
superDict[refID].append(int(line[-1]))
#End loop through
#Make table by gene
overallGeneList = utils.uniquify(overallGeneList)
#use enhancer rank to order
rankOrder = utils.order([min(rankDict[x]) for x in overallGeneList])
usedNames = []
for i in rankOrder:
refID = overallGeneList[i]
geneName = startDict[refID]['name']
if usedNames.count(geneName) > 0 and uniqueGenes == True:
continue
else:
usedNames.append(geneName)
proxEnhancers = geneDict['overlapping'][refID]+geneDict['proximal'][refID]
superStatus = max(superDict[refID])
enhancerRanks = join([str(x) for x in rankDict[refID]],',')
newLine = [geneName,refID,join(proxEnhancers,','),enhancerRanks,superStatus]
geneToEnhancerTable.append(newLine)
#resort enhancerToGeneTable
if noFormatTable:
return enhancerToGeneTable,geneToEnhancerTable
else:
enhancerOrder = utils.order([int(line[-2]) for line in enhancerToGeneTable[1:]])
sortedTable = [enhancerToGeneTable[0]]
for i in enhancerOrder:
sortedTable.append(enhancerToGeneTable[(i+1)])
return sortedTable,geneToEnhancerTable
def mapBamToGFF(bamFile,
gff,
sense='.',
extension=200,
rpm=False,
clusterGram=None,
matrix=None):
'''maps reads from a bam to a gff'''
#creating a new gff to output
newGFF = []
#reading in the bam
bam = utils.Bam(bamFile)
#getting RPM normalization
if rpm:
MMR = round(float(bam.getTotalReads('mapped')) / 1000000, 4)
else:
MMR = 1
print('using a MMR value of %s' % (MMR))
#creating a sense trans
senseTrans = string.maketrans('-+.', '+-+')
#reading in the gff
if type(gff) == str:
gff = utils.parseTable(gff, '\t')
#setting up a clustergram table
if clusterGram:
binSize = int(clusterGram)
binSizeList = []
#now go through each line of the gff and make sure they're all the same length
for i in range(0, len(gff), 1):
line = gff[i]
gffLocus = utils.Locus(line[0], int(line[3]), int(line[4]),
line[6], line[1])
binSizeList.append(gffLocus.len() / binSize)
binSizeList = utils.uniquify(binSizeList)
if len(binSizeList) > 1:
print(
'WARNING: lines in gff are of different length. Output clustergram will have variable row length'
)
newGFF.append(['GENE_ID', 'locusLine'] + [
str(x * binSize) + '_' + bamFile.split('/')[-1]
for x in range(1,
max(binSizeList) + 1, 1)
])
#setting up a maxtrix table
if matrix:
newGFF.append(['GENE_ID', 'locusLine'] + [
'bin_' + str(n) + '_' + bamFile.split('/')[-1]
for n in range(1,
int(matrix) + 1, 1)
])
nBin = int(matrix)
# Try to use the bamliquidatior script on cluster, otherwise, failover to local (in path), otherwise fail.
bamliquidatorString = '/usr/bin/bamliquidator'
if not os.path.isfile(bamliquidatorString):
bamliquidatorString = './bamliquidator'
if not os.path.isfile(bamliquidatorString):
raise ValueError('bamliquidator not found in path')
#getting and processing reads for gff lines
ticker = 0
print('Number lines processed')
for line in gff:
line = line[0:9]
if ticker % 100 == 0:
print(ticker)
ticker += 1
gffLocus = utils.Locus(line[0], int(line[3]), int(line[4]), line[6],
line[1])
#get the nBin and binSize
if clusterGram:
nBin = gffLocus.len() / int(clusterGram)
binSize = int(clusterGram)
if matrix:
nBin = int(matrix)
binSize = gffLocus.len() / nBin
#some regions will be too short to get info on
if binSize == 0:
clusterLine = [gffLocus.ID(),
gffLocus.__str__()] + ['NA'] * nBin
newGFF.append(clusterLine)
continue
#flippy flip if sense is negative
if sense == '-':
bamSense = string.translate(gffLocus.sense(), senseTrans)
elif sense == '+':
bamSense = gffLocus.sense()
else:
bamSense = '.'
#using the bamLiquidator to get the readstring
#print('using nBin of %s' % nBin)
bamCommand = "%s %s %s %s %s %s %s %s" % (
bamliquidatorString, bamFile, line[0], gffLocus.start(),
gffLocus.end(), bamSense, nBin, extension)
#print(bamCommand)
getReads = subprocess.Popen(bamCommand,
stdin=subprocess.PIPE,
stderr=subprocess.PIPE,
stdout=subprocess.PIPE,
shell=True)
readString, stderr = getReads.communicate()
if stderr:
print("STDERR out: %s" % (stderr))
denList = readString.split('\n')[:-1]
#print("denlist is: %s" % denList)
#flip the denList if the actual gff region is -
if gffLocus.sense() == '-':
denList = denList[::-1]
#converting from units of total bp of read sequence per bin to rpm/bp
denList = [round(float(x) / binSize / MMR, 4) for x in denList]
#if the gff region is - strand, flip the
clusterLine = [gffLocus.ID(), gffLocus.__str__()] + denList
newGFF.append(clusterLine)
return newGFF
def findCanidateTFs(annotationFile, superLoci, expressedNM, TFlist, refseqToNameDict, projectFolder, projectName):
'''
find all TFs within 1Mb of the super-enhancer center that are considered expressed
return a dictionary keyed by TF that points to a list of super-enhancer loci
'''
print 'FINDING CANIDATE TFs'
startDict = utils.makeStartDict(annotationFile)
# Find the location of the TSS of all transcripts (NMid) considered expressed
tssLoci = []
for geneID in expressedNM:
tssLoci.append(utils.makeTSSLocus(geneID,startDict,0,0))
tssCollection = utils.LocusCollection(tssLoci,50)
# Assign all transcripts (NMid) that are TFs to a super-enhancer if it is the closest gene
seAssignment = []
seAssignmentGene = []
TFandSuperDict = {}
for superEnh in superLoci:
seCenter = (superEnh.start() + superEnh.end()) / 2
# Find all transcripts whose TSS occur within 1Mb of the SE center
searchLocus = utils.Locus(superEnh.chr(), superEnh.start()-1000000, superEnh.end()+1000000, '.')
allEnhancerLoci = tssCollection.getOverlap(searchLocus)
allEnhancerGenes = [locus.ID() for locus in allEnhancerLoci]
# Find the transcript that is closest to the center
if allEnhancerGenes:
distList = [abs(seCenter - startDict[geneID]['start'][0]) for geneID in allEnhancerGenes]
closestGene = allEnhancerGenes[distList.index(min(distList))]
else:
closestGene = ''
seAssignment.append([superEnh.chr(), superEnh.start(), superEnh.end(), closestGene])
# Select the transcript if it is a TF, and allow for a TF to have multiple SEs
if closestGene in TFlist and closestGene not in TFandSuperDict.keys():
TFandSuperDict[closestGene] = [superEnh]
elif closestGene in TFlist and closestGene in TFandSuperDict.keys():
TFandSuperDict[closestGene].append(superEnh)
# Convert the selected TF NMids to gene names
if closestGene != '':
geneName = refseqToNameDict[closestGene]
seAssignmentGene.append([superEnh.chr(), superEnh.start(), superEnh.end(), geneName])
# Output the list of SE-assigned transcripts (NMids)
seAssignmentFile = projectFolder + projectName + '_SE_ASSIGNMENT_TRANSCRIPT.txt'
utils.unParseTable(seAssignment, seAssignmentFile, '\t')
# Output the list of SE-assigned genes
seAssignmentGeneFile = projectFolder + projectName + '_SE_ASSIGNMENT_GENE.txt'
utils.unParseTable(seAssignmentGene, seAssignmentGeneFile, '\t')
print 'Number of canidate TFs:', len(TFandSuperDict)
return TFandSuperDict
def rank_eboxes(nb_all_chip_dataFile,mycn_gff_path,macsFolder,genomeDirectory,window = 100):
'''
uses the conserved MYCN sites and ranks eboxes within them
by average background subtracted signal
searches 100bp (window variable) from mycn summits
'''
window = int(window)
#bring in the conserved mycn region
print('making gff of nb mycn summits')
nb_mycn_gff = utils.parseTable(mycn_gff_path,'\t')
nb_mycn_collection = utils.gffToLocusCollection(nb_mycn_gff,50)
dataDict =pipeline_dfci.loadDataTable(nb_all_chip_dataFile)
names_list = [name for name in dataDict.keys() if name.count('MYCN') == 1]
names_list.sort()
summit_loci = []
#first makes a gff of all summits +/- 100bp for all nb mycn datasets
for name in names_list:
summit_bed_path = '%s%s/%s_summits.bed' % (macsFolder,name,name)
summit_bed = utils.parseTable(summit_bed_path,'\t')
for line in summit_bed:
summit_locus = utils.Locus(line[0],int(line[1])-window,int(line[2])+window,'.',line[3])
if len(nb_mycn_collection.getOverlap(summit_locus)) > 0:
summit_loci.append(summit_locus)
summit_collection =utils.LocusCollection(summit_loci,50)
summit_merged_collection = summit_collection.stitchCollection()
summit_gff = utils.locusCollectionToGFF(summit_merged_collection)
summit_gff_path = '%sHG19_NB_MYCN_SUMMITS_-%s_+%s.gff' % (gffFolder,window,window)
utils.unParseTable(summit_gff,summit_gff_path,'\t')
#this is borrowed from above and maps chip-seq signal to the gff
print('mapping to nb mycn summits and making signal dict')
gffList = [summit_gff_path]
summit_signal_path = pipeline_dfci.map_regions(nb_all_chip_dataFile,gffList)
mycnSignalTable = utils.parseTable(summit_signal_path,'\t')
#making a signal dictionary for MYCN binding
names_list = ['BE2C_MYCN','KELLY_MYCN','NGP_MYCN','SHEP21_0HR_MYCN_NOSPIKE']
background_list = [dataDict[name]['background'] for name in names_list]
header = mycnSignalTable[0]
chip_columns = [header.index(name) for name in names_list]
background_columns = [header.index(background_name) for background_name in background_list]
mycn_sig_dict = {}
for line in mycnSignalTable[1:]:
line_sig = []
for i in range(len(names_list)):
line_sig.append(float(line[chip_columns[i]]) - float(line[background_columns[i]]))
region_id = line[1]
coords = [int(x) for x in line[1].split(':')[-1].split('-')]
line_length = coords[1]-coords[0]
mycn_sig_dict[region_id] = numpy.mean(line_sig)*line_length
#now for each region find the eboxes and then add up the signal
print('making ebox ranking')
ebox_list = ['CACGTG','CAGTTG','CAAGTG','CAGGTG','CAATTG','CAAATG','CATCTG','CAGCTG','CATGTG','CATATG']
eboxDict = {}
for ebox in ebox_list:
eboxDict[ebox] = []
ticker = 0
for line in summit_gff:
if ticker % 1000 == 0:
print(ticker)
ticker+=1
chrom = line[0]
sense = '.'
start = int(line[3])
end = int(line[4])
region_id = '%s(%s):%s-%s' % (line[0],line[6],line[3],line[4])
signal = mycn_sig_dict[region_id]
sequenceLine = utils.fetchSeq(genomeDirectory,chrom,start,end,True)
motifVector = []
matches = re.finditer('CA..TG',str.upper(sequenceLine))
if matches:
for match in matches:
motifVector.append(match.group())
#count only 1 of each motif type per line
#motifVector = utils.uniquify(motifVector)
for motif in motifVector:
if ebox_list.count(motif) > 0:
eboxDict[motif].append(signal)
else:
eboxDict[utils.revComp(motif)].append(signal)
eboxTable =[]
eboxTableOrdered =[['EBOX','OCCURENCES','AVG_HEIGHT']]
for ebox in eboxDict.keys():
newLine = [ebox,len(eboxDict[ebox]),numpy.mean(eboxDict[ebox])]
eboxTable.append(newLine)
occurenceOrder = utils.order([line[2] for line in eboxTable],decreasing=True)
for x in occurenceOrder:
eboxTableOrdered.append(eboxTable[x])
print(eboxTableOrdered)
ebox_outfile = '%sHG19_NB_MYCN_CONSERVED_SUMMITS_-%s_+%s_EBOX_RANK.txt' % (tableFolder,window,window)
utils.unParseTable(eboxTableOrdered,ebox_outfile,'\t')
return ebox_outfile
def collapseFimo(fimo_output, gene_to_enhancer_dict, candidate_tf_list,
output_folder, analysis_name, motifConvertFile):
'''
collapses motifs from fimo
for each source node (TF) and each target node (gene enhancer regions), collapse motif instances
then spit out a ginormous set of beds and a single crazy collapsed bed
'''
#first build up the motif name conversion database
motifDatabase = utils.parseTable(motifConvertFile, '\t')
motifDatabaseDict = defaultdict(list)
# The reverse of the other dict, from motif name to gene name
# a motif can go to multiple genes
for line in motifDatabase:
motifDatabaseDict[line[0]].append(line[1])
#make the folder to store motif beds
utils.formatFolder('%smotif_beds/' % (output_folder), True)
edgeDict = {}
#first layer are source nodes
for tf in candidate_tf_list:
edgeDict[tf] = defaultdict(
list
) #next layer are target nodes which are derived from the fimo output
fimoTable = utils.parseTable(fimo_output, '\t')
print(fimo_output)
#fimo sometimes puts the region in either the first or second column
fimo_line = fimoTable[1]
if fimo_line[1].count('|') > 0:
region_index = 1
else:
region_index = 2
print('USING COLUMN %s OF FIMO OUTPUT FOR REGION' % (region_index))
for line in fimoTable[1:]:
source_tfs = motifDatabaseDict[line[0]] #motifId
for source in source_tfs:
if candidate_tf_list.count(source) == 0:
continue
region = line[region_index].split('|')
target = region[0]
if region_index == 2:
target_locus = utils.Locus(region[1],
int(region[2]) + int(line[3]),
int(region[2]) + int(line[4]), '.')
else:
target_locus = utils.Locus(region[1],
int(region[2]) + int(line[2]),
int(region[2]) + int(line[3]), '.')
#what's missing here is the enhancer id of the target locus
try:
edgeDict[source][target].append(target_locus)
except KeyError:
print('this motif is not in the network')
print(line)
sys.exit()
#now we actually want to collapse this down in a meaningful way
#overlapping motifs count as a single binding site. This way a TF with tons of motifs
#that finds the same site over and over again doesn't get over counted
all_bed = []
all_bed_path = '%s%s_all_motifs.bed' % (output_folder, analysis_name)
for tf in candidate_tf_list:
print(tf)
target_nodes = edgeDict[tf].keys()
bed_header = [
'track name = "%s" description="%s motifs in %s"' %
(tf, tf, analysis_name)
]
all_bed.append(bed_header)
target_bed = [bed_header]
target_bed_path = '%smotif_beds/%s_motifs.bed' % (output_folder, tf)
for target in target_nodes:
edgeCollection = utils.LocusCollection(edgeDict[tf][target], 50)
edgeCollection = edgeCollection.stitchCollection()
edgeLoci = edgeCollection.getLoci()
edgeDict[tf][target] = edgeLoci
for locus in edgeLoci:
bed_line = [
locus.chr(),
locus.start(),
locus.end(), target, '', '+'
]
target_bed.append(bed_line)
all_bed.append(bed_line)
utils.unParseTable(target_bed, target_bed_path, '\t')
#now the loci are all stitched up
utils.unParseTable(all_bed, all_bed_path, '\t')
return edgeDict
def make_shep_on_mycn_landscape(shep_on_dataFile):
'''
finds mycn peaks in shep21 that are conserved in nb and segregates them into promoter or enhancer
'''
dataDict = pipeline_dfci.loadDataTable(shep_on_dataFile)
print('LOADING SHEP ON MYCN SITES')
#load all of the shep_on sites
# shep_on_gff_path = '%smeta_rose/SHEP_ON_MYC/gff/HG19_SHEP_ON_MYC_ALL_-0_+0.gff' % (projectFolder)
# shep_on_gff = utils.parseTable(shep_on_gff_path,'\t')
shep_on_bed_path = '%sSHEP_6HR_MYCN_peaks.bed' % (macsEnrichedFolder)
shep_on_bed = utils.parseTable(shep_on_bed_path,'\t')
shep_on_gff = utils.bedToGFF(shep_on_bed)
#now get the conserved NB MYCN regions
nb_conserved_mycn_gff_file = '%sHG19_NB_MYCN_CONSERVED_-0_+0.gff' % (gffFolder)
nb_conserved_mycn_collection = utils.gffToLocusCollection(nb_conserved_mycn_gff_file)
print('LOADING SHEP ACTIVE ENHANCERS')
#make a collection of enhancers
shep_enhancer_file = '%smeta_rose/SHEP_ON_H3K27AC/SHEP_ON_H3K27AC_AllEnhancers.table.txt' % (projectFolder)
shep_enhancer_collection = utils.makeSECollection(shep_enhancer_file,'SHEP_H3K27AC')
#now get the active promoters
print('LOADING SHEP ACTIVE PROMOTERS')
startDict = utils.makeStartDict(annotFile)
shep_transcribed_file = '%sHG19_SHEP_ON_H3K27AC_ACTIVE.txt' % (geneListFolder)
shep_transcribed_table = utils.parseTable(shep_transcribed_file,'\t')
transcribedList = [line[1] for line in shep_transcribed_table]
tssLoci = []
for refID in transcribedList:
tssLoci.append(utils.makeTSSLocus(refID,startDict,1000,1000))
shep_tss_collection = utils.LocusCollection(tssLoci,50)
#now initialize the 6 gffs we will need
shep_mycn_gff = []
shep_mycn_gff_5kb = []
shep_mycn_gff_1kb = []
shep_mycn_promoter_gff = []
shep_mycn_promoter_gff_1kb = []
shep_mycn_promoter_gff_5kb = []
shep_mycn_enhancer_gff = []
shep_mycn_enhancer_gff_1kb = []
shep_mycn_enhancer_gff_5kb = []
#and their respective file names
shep_mycn_gff_file = '%sHG19_SHEP_MYCN_CONSERVED_-0_+0.gff' % (gffFolder)
shep_mycn_gff_5kb_file = '%sHG19_SHEP_MYCN_CONSERVED_-5kb_+5kb.gff' % (gffFolder)
shep_mycn_gff_1kb_file = '%sHG19_SHEP_MYCN_CONSERVED_-1kb_+1kb.gff' % (gffFolder)
shep_mycn_promoter_gff_file = '%sHG19_SHEP_MYCN_CONSERVED_PROMOTER_-0_+0.gff' % (gffFolder)
shep_mycn_promoter_gff_5kb_file = '%sHG19_SHEP_MYCN_CONSERVED_PROMOTER_-5kb_+5kb.gff' % (gffFolder)
shep_mycn_promoter_gff_1kb_file = '%sHG19_SHEP_MYCN_CONSERVED_PROMOTER_-1kb_+1kb.gff' % (gffFolder)
shep_mycn_enhancer_gff_file = '%sHG19_SHEP_MYCN_CONSERVED_ENHANCER_-0_+0.gff' % (gffFolder)
shep_mycn_enhancer_gff_5kb_file = '%sHG19_SHEP_MYCN_CONSERVED_ENHANCER_-5kb_+5kb.gff' % (gffFolder)
shep_mycn_enhancer_gff_1kb_file = '%sHG19_SHEP_MYCN_CONSERVED_ENHANCER_-1kb_+1kb.gff' % (gffFolder)
print('ITERATING THROUGH SHEP MYCN PEAKS')
ticker = 0
enhancer = 0
promoter = 0
other = 0
for line in shep_on_gff:
if ticker % 1000 == 0:
print ticker
ticker+=1
peakID = '%s_%s' % ('SHEP_MYCN',str(ticker))
lineLocus = utils.Locus(line[0],line[3],line[4],'.',peakID)
if nb_conserved_mycn_collection.getOverlap(lineLocus):
gffLine = [line[0],peakID,peakID,line[3],line[4],'','.','',peakID]
peakCenter = (int(line[3]) + int(line[4]))/2
gffLine_5kb = [line[0],peakID,peakID,peakCenter - 5000,peakCenter + 5000,'','.','',peakID]
#the 1kb is not a center +/- but a flank
gffLine_1kb = [line[0],peakID,peakID,int(line[3]) - 1000,int(line[4]) + 1000,'','.','',peakID]
shep_mycn_gff.append(gffLine)
shep_mycn_gff_5kb.append(gffLine_5kb)
shep_mycn_gff_1kb.append(gffLine_1kb)
#tss overlap should take precedence over enhancer overlap
if shep_tss_collection.getOverlap(lineLocus,'both'):
shep_mycn_promoter_gff.append(gffLine)
shep_mycn_promoter_gff_5kb.append(gffLine_5kb)
shep_mycn_promoter_gff_1kb.append(gffLine_1kb)
promoter+=1
#now check for enhancer overlap
elif shep_enhancer_collection.getOverlap(lineLocus,'both'):
shep_mycn_enhancer_gff.append(gffLine)
shep_mycn_enhancer_gff_5kb.append(gffLine_5kb)
shep_mycn_enhancer_gff_1kb.append(gffLine_1kb)
enhancer+=1
else:
other+=1
print('Of %s shep on mycn peaks' % (len(shep_on_gff)))
print('%s are promoter' % (promoter))
print('%s are enhancer' % (enhancer))
print('%s are other' % (other))
#now write out the gffs
utils.unParseTable(shep_mycn_gff,shep_mycn_gff_file,'\t')
utils.unParseTable(shep_mycn_gff_5kb,shep_mycn_gff_5kb_file,'\t')
utils.unParseTable(shep_mycn_gff_1kb,shep_mycn_gff_1kb_file,'\t')
utils.unParseTable(shep_mycn_promoter_gff,shep_mycn_promoter_gff_file,'\t')
utils.unParseTable(shep_mycn_promoter_gff_5kb,shep_mycn_promoter_gff_5kb_file,'\t')
utils.unParseTable(shep_mycn_promoter_gff_1kb,shep_mycn_promoter_gff_1kb_file,'\t')
utils.unParseTable(shep_mycn_enhancer_gff,shep_mycn_enhancer_gff_file,'\t')
utils.unParseTable(shep_mycn_enhancer_gff_5kb,shep_mycn_enhancer_gff_5kb_file,'\t')
utils.unParseTable(shep_mycn_enhancer_gff_1kb,shep_mycn_enhancer_gff_1kb_file,'\t')
# First, load in the node TFs, ATAC peaks and super enhancer regions we'll consider for this analysis
# From networks already constructed from CRC2.py
node_file = '/crusader/projects/cll/final/network/lines/zinba/' + projectName + '/' + projectName + '_NODELIST.txt'
node_table = utils.parseTable(node_file, '\t')
nodelist = [x[0] for x in node_table]
print nodelist
super_enhancer_file = '/crusader/projects/cll/final/rose/' + projectName + '_H3K27ac/' + projectName + '_H3K27ac_peaks_SuperEnhancers.table.txt'
se_table = utils.parseTable(super_enhancer_file, '\t')
subpeak_file = '/crusader/projects/cll/final/zinba/lines/MEC1_ATAC/MEC1_ATAC.peaks.bed'
subpeak_table = utils.parseTable(subpeak_file, '\t')
subpeak_loci = []
for line in subpeak_table:
subpeak_loci.append(utils.Locus(line[0], line[1], line[2], '.'))
subpeak_collection = utils.LocusCollection(subpeak_loci, 100)
subpeak_dict = {} # key is enhancer ID, points to a list of loci
# assign subpeak Loci to each super enhancer
fasta = []
se_namelist = []
for line in se_table[6:]:
se_id = line[0]
se_namelist.append(se_id)
subpeak_dict[se_id] = []
se_locus = utils.Locus(line[1], line[2], line[3], '.')
overlaps = subpeak_collection.getOverlap(se_locus)
def main():
projectFolder = '/storage/goodell/home/jmreyes/projects/amish_ayala/'
#gather up DMR tables
#ayala MUT vs WT
mutWT_hypo = utils.parseTable(projectFolder + 'bed/hypoDMRsWT.vs.Mut.bed',
'\t')
mutWT_hyper = utils.parseTable(
projectFolder + 'bed/hyperDMRsWT.vs.Mut.bed', '\t')
mutWT_control = utils.parseTable(
projectFolder + 'bed/Control_nonDMRsWT.vs.Mut.bed', '\t')
#ley all
tbrs_all = utils.parseTable(projectFolder + 'bed/TBRS_DMRs.bed', '\t')
aml_all = utils.parseTable(projectFolder + 'bed/AML_DMRs.bed', '\t')
tbrs_hypo = []
tbrs_hyper = []
aml_hypo = []
aml_hyper = []
tbrs_all_loci = []
aml_all_loci = []
for line in tbrs_all:
chrom = 'chr' + line[0]
start = line[1]
end = line[2]
if 'hypo' in line:
tbrs_all_loci.append(
utils.Locus(
chrom, start, end, '.', 'tbrs_all_hypo_' + str(chrom) +
':' + str(start) + '-' + str(end)))
elif 'hyper' in line:
tbrs_all_loci.append(
utils.Locus(
chrom, start, end, '.', 'tbrs_all_hyper_' + str(chrom) +
':' + str(start) + '-' + str(end)))
for line in aml_all:
chrom = 'chr' + line[0]
start = line[1]
end = line[2]
if 'hypo' in line:
aml_all_loci.append(
utils.Locus(
chrom, start, end, '.', 'aml_all_hypo_' + str(chrom) +
':' + str(start) + '-' + str(end)))
elif 'hyper' in line:
aml_all_loci.append(
utils.Locus(
chrom, start, end, '.', 'aml_all_hyper_' + str(chrom) +
':' + str(start) + '-' + str(end)))
mutWT_hypo_loci = []
for line in mutWT_hypo:
chrom = line[0]
start = line[1]
end = line[2]
sense = '.'
locusID = 'hypo_' + str(chrom) + ':' + str(start) + '-' + str(end)
new_line = utils.Locus(chrom, start, end, '.', locusID)
mutWT_hypo_loci.append(new_line)
mutWT_hyper_loci = []
for line in mutWT_hyper:
chrom = line[0]
start = line[1]
end = line[2]
sense = '.'
locusID = 'hyper_' + str(chrom) + ':' + str(start) + '-' + str(end)
new_line = utils.Locus(chrom, start, end, '.', locusID)
mutWT_hyper_loci.append(new_line)
print len(mutWT_hyper_loci)
print len(mutWT_hypo_loci)
mutWT_all_loci = mutWT_hyper_loci + mutWT_hypo_loci
mutWT_hypo_LC = utils.LocusCollection(mutWT_hypo_loci)
tbrs_all_LC = utils.LocusCollection(tbrs_all_loci)
aml_all_LC = utils.LocusCollection(aml_all_loci)
tbrs_all_overlap = []
aml_all_overlap = []
for locus in mutWT_hypo_LC.getLoci():
tbrs_overlap = tbrs_all_LC.getOverlap(locus, 'both')
if len(tbrs_overlap) > 0:
for overlapLocus in tbrs_overlap:
overlapChrom = overlapLocus.chr()
overlapStart = overlapLocus.start()
overlapEnd = overlapLocus.end()
tbrs_all_overlap.append([
locus.ID(), overlapChrom, overlapStart, overlapEnd,
overlapLocus.ID()
])
aml_overlap = aml_all_LC.getOverlap(locus, 'both')
if len(aml_overlap) > 0:
for overlapLocus in aml_overlap:
overlapChrom = overlapLocus.chr()
overlapStart = overlapLocus.start()
overlapEnd = overlapLocus.end()
aml_all_overlap.append([
locus.ID(), overlapChrom, overlapStart, overlapEnd,
overlapLocus.ID()
])
utils.unParseTable(tbrs_all_overlap,
projectFolder + 'tables/DMRsvsTBRS_all_overlaps.txt',
'\t')
utils.unParseTable(aml_all_overlap,
projectFolder + 'tables/DMRsvsAML_all_overlaps.txt',
'\t')
def mapGFFLineToAnnot(gffLine,
outFolder,
nBins,
geneDict,
txCollection,
sense='both',
header=''):
'''
for every line produces a file with all of the rectangles to draw
'''
if len(header) == 0:
gffString = '%s_%s_%s_%s' % (gffLine[0], gffLine[6], gffLine[3],
gffLine[4])
else:
gffString = header
diagramTable = [[0, 0, 0, 0]]
nameTable = [['', 0, 0]]
gffLocus = utils.Locus(gffLine[0], int(gffLine[3]), int(gffLine[4]),
gffLine[6], gffLine[1])
scaleFactor = float(nBins) / gffLocus.len()
# plotting buffer for diagrams
plotBuffer = int(gffLocus.len() / float(nBins) * 20)
overlapLoci = txCollection.getOverlap(gffLocus, sense='both')
geneList = [locus.ID() for locus in overlapLoci]
if gffLine[6] == '-':
refPoint = int(gffLine[4])
else:
refPoint = int(gffLine[3])
offsetCollection = utils.LocusCollection([], 500)
for geneID in geneList:
gene = geneDict[geneID]
print(gene.commonName())
if len(gene.commonName()) > 1:
name = gene.commonName()
else:
name = geneID
offset = 4 * len(offsetCollection.getOverlap(gene.txLocus()))
offsetCollection.append(
utils.makeSearchLocus(gene.txLocus(), plotBuffer, plotBuffer))
# write the name of the gene down
if gene.sense() == '+':
geneStart = gene.txLocus().start()
else:
geneStart = gene.txLocus().end()
geneStart = abs(geneStart - refPoint) * scaleFactor
nameTable.append([name, geneStart, -2 - offset])
# draw a line across the entire txLocus
[start, stop] = [
abs(x - refPoint) * scaleFactor for x in gene.txLocus().coords()
]
diagramTable.append([start, -0.01 - offset, stop, 0.01 - offset])
# now draw thin boxes for all txExons
if len(gene.txExons()) > 0:
for txExon in gene.txExons():
[start, stop] = [
abs(x - refPoint) * scaleFactor for x in txExon.coords()
]
diagramTable.append([start, -0.5 - offset, stop, 0.5 - offset])
# now draw fatty boxes for the coding exons if any
if len(gene.cdExons()) > 0:
for cdExon in gene.cdExons():
[start, stop] = [
abs(x - refPoint) * scaleFactor for x in cdExon.coords()
]
diagramTable.append([start, -1 - offset, stop, 1 - offset])
utils.unParseTable(diagramTable,
outFolder + gffString + '_diagramTemp.txt', '\t')
utils.unParseTable(nameTable, outFolder + gffString + '_nameTemp.txt',
'\t')
def geneToEnhancerDict(genome, enhancer_file, activity_path):
'''
Assign each Super-Enhancer to the closest active TSS to its center
Return a dictionary keyed by TF that points to a list of loci
'''
print('Identifying enhancers and target genes from %s' % (enhancer_file))
#should this do gene assignment????
#for now assume gene assignment has been done
#can later toggle to do gene assignment
#first load the TF lists
tf_table = utils.parseTable(genome.returnFeature('tf_file'), '\t')
motif_table = utils.parseTable(genome.returnFeature('motif_convert'), '\t')
#this gives all tfs that have a motif
motif_tfs = utils.uniquify([line[1] for line in motif_table])
#intersect w/ the activity table
if len(activity_path) > 0:
activity_table = utils.parseTable(activity_path, '\t')
#figure out the right column for actual gene names (basically not NM or NR and not a numeral)
for i in range(len(activity_table[0])):
# try:
# foo = int(activity_table[0][i])
# except ValueError: # case where it is not an integer
if activity_table[0][i][0:2] != 'NM' and activity_table[0][i][
0:2] != 'NR': #assumes refseq
gene_col = i
break
print('using column %s of %s gene activity table for common names' %
(gene_col + 1, activity_path))
active_gene_list = [
string.upper(line[gene_col]) for line in activity_table
]
tf_list_refseq = [
line[0] for line in tf_table if active_gene_list.count(line[1]) > 0
and motif_tfs.count(line[1]) > 0
]
tf_list_name = utils.uniquify([
line[1] for line in tf_table if active_gene_list.count(line[1]) > 0
and motif_tfs.count(line[1]) > 0
])
else:
tf_list_refseq = [
line[0] for line in tf_table if motif_tfs.count(line[1]) > 0
]
tf_list_name = [
line[1] for line in tf_table if motif_tfs.count(line[1]) > 0
]
print('Identified %s TFs from %s that have motifs' %
(len(tf_list_name), genome.returnFeature('tf_file')))
#keyed by gene with loci objects in the list
gene_to_enhancer_dict = defaultdict(list)
enhancer_to_gene_dict = defaultdict(list)
#assuming id,chrom,start,stop w/ gene names in the last 3 columns per standard ROSE output
enhancer_table = utils.parseTable(enhancer_file, '\t')
print('Analyzing %s cis-regulatory regions' % (len(enhancer_table)))
#now let's make the enhancer table by region and then by gene
enhancerTable = [['ENHANCER_ID', 'CHROM', 'START', 'STOP', 'GENE_LIST']]
enhancerTFTable = [['ENHANCER_ID', 'CHROM', 'START', 'STOP', 'GENE_LIST']]
geneTable = [['GENE', 'TF', 'CHROM', 'START', 'STOP', 'ENHANCER_ID']]
geneTFTable = [['GENE', 'CHROM', 'START', 'STOP', 'ENHANCER_ID']]
geneSummaryTable = [['GENE', 'TF', 'ENHANCER_LIST']]
#will need to track which ones are TFs
candidate_tf_list = []
#find the columns for gene assignment
header = enhancer_table[0]
header_length = len(enhancer_table[0])
closest_index = header.index('CLOSEST_GENE')
proximal_index = header.index('PROXIMAL_GENES')
overlap_index = header.index('OVERLAP_GENES')
for line in enhancer_table[1:]:
if len(
line
) != header_length: #don't bother trying to figure out lines w/o target genes
continue
enhancer_locus = utils.Locus(line[1], line[2], line[3], '.', line[0])
closest_gene_list = line[closest_index].split(',')
proximal_gene_list = line[proximal_index].split(',')
overlap_gene_list = line[overlap_index].split(',')
all_gene_list = closest_gene_list + proximal_gene_list + overlap_gene_list
all_gene_list = [string.upper(gene) for gene in all_gene_list]
#print(all_gene_list)
#print(activity_path)
#print(active_gene_list)
#gets a unique list of all tfs
if len(activity_path) > 0:
all_gene_list = utils.uniquify([
gene for gene in all_gene_list
if active_gene_list.count(gene) > 0
])
else:
all_gene_list = utils.uniquify(all_gene_list)
candidate_gene_list = utils.uniquify(
[gene for gene in all_gene_list if tf_list_name.count(gene) > 0])
if len(all_gene_list) > 0:
for gene in all_gene_list:
gene_to_enhancer_dict[gene].append(enhancer_locus)
enhancer_to_gene_dict[enhancer_locus].append(gene)
newLine = line[0:4] + [','.join(all_gene_list)]
else:
newLine = line[0:4] + ['']
enhancerTable.append(newLine)
if len(candidate_gene_list) > 0:
tfLine = line[0:4] + [','.join(candidate_gene_list)]
enhancerTFTable.append(tfLine)
#now iterate through each gene and list the enhancers
gene_list = gene_to_enhancer_dict.keys()
print(gene_list)
gene_list.sort()
for gene in gene_list:
if tf_list_name.count(gene) > 0:
tf_status = 1
candidate_tf_list.append(gene)
else:
tf_status = 0
enhancer_loci = gene_to_enhancer_dict[gene]
enhancerString = ','.join(
[enhancer.ID() for enhancer in enhancer_loci])
geneSummaryTable.append([gene, tf_status, enhancerString])
for enhancer in enhancer_loci:
newLine = [
gene, tf_status,
enhancer.chr(),
enhancer.start(),
enhancer.end(),
enhancer.ID()
]
geneTable.append(newLine)
if tf_status == 1:
newLine = [
gene,
enhancer.chr(),
enhancer.start(),
enhancer.end(),
enhancer.ID()
]
geneTFTable.append(newLine)
return geneTable, geneTFTable, enhancerTable, enhancerTFTable, geneSummaryTable, candidate_tf_list, gene_to_enhancer_dict
def mapEnhancerToGeneTop(rankByBamFile, controlBamFile, genome, annotFile, enhancerFile, transcribedFile='', uniqueGenes=True, searchWindow=50000, noFormatTable=False):
'''
maps genes to enhancers. if uniqueGenes, reduces to gene name only. Otherwise, gives for each refseq
'''
startDict = utils.makeStartDict(annotFile)
enhancerName = enhancerFile.split('/')[-1].split('.')[0]
enhancerTable = utils.parseTable(enhancerFile, '\t')
# internal parameter for debugging
byRefseq = False
if len(transcribedFile) > 0:
transcribedTable = utils.parseTable(transcribedFile, '\t')
transcribedGenes = [line[1] for line in transcribedTable]
else:
transcribedGenes = startDict.keys()
print('MAKING TRANSCRIPT COLLECTION')
transcribedCollection = utils.makeTranscriptCollection(
annotFile, 0, 0, 500, transcribedGenes)
print('MAKING TSS COLLECTION')
tssLoci = []
for geneID in transcribedGenes:
tssLoci.append(utils.makeTSSLocus(geneID, startDict, 0, 0))
# this turns the tssLoci list into a LocusCollection
# 50 is the internal parameter for LocusCollection and doesn't really
# matter
tssCollection = utils.LocusCollection(tssLoci, 50)
geneDict = {'overlapping': defaultdict(
list), 'proximal': defaultdict(list)}
# dictionaries to hold ranks and superstatus of gene nearby enhancers
rankDict = defaultdict(list)
superDict = defaultdict(list)
# list of all genes that appear in this analysis
overallGeneList = []
# find the damn header
for line in enhancerTable:
if line[0][0] == '#':
continue
else:
header = line
break
if noFormatTable:
# set up the output tables
# first by enhancer
enhancerToGeneTable = [
header + ['OVERLAP_GENES', 'PROXIMAL_GENES', 'CLOSEST_GENE']]
else:
# set up the output tables
# first by enhancer
enhancerToGeneTable = [
header[0:9] + ['OVERLAP_GENES', 'PROXIMAL_GENES', 'CLOSEST_GENE'] + header[-2:]]
# next by gene
geneToEnhancerTable = [
['GENE_NAME', 'REFSEQ_ID', 'PROXIMAL_ENHANCERS']]
# next make the gene to enhancer table
geneToEnhancerTable = [
['GENE_NAME', 'REFSEQ_ID', 'PROXIMAL_ENHANCERS', 'ENHANCER_RANKS', 'IS_SUPER', 'ENHANCER_SIGNAL']]
for line in enhancerTable:
if line[0][0] == '#' or line[0][0] == 'R':
continue
enhancerString = '%s:%s-%s' % (line[1], line[2], line[3])
enhancerLocus = utils.Locus(line[1], line[2], line[3], '.', line[0])
# overlapping genes are transcribed genes whose transcript is directly
# in the stitchedLocus
overlappingLoci = transcribedCollection.getOverlap(
enhancerLocus, 'both')
overlappingGenes = []
for overlapLocus in overlappingLoci:
overlappingGenes.append(overlapLocus.ID())
# proximalGenes are transcribed genes where the tss is within 50kb of
# the boundary of the stitched loci
proximalLoci = tssCollection.getOverlap(
utils.makeSearchLocus(enhancerLocus, searchWindow, searchWindow), 'both')
proximalGenes = []
for proxLocus in proximalLoci:
proximalGenes.append(proxLocus.ID())
distalLoci = tssCollection.getOverlap(
utils.makeSearchLocus(enhancerLocus, 1000000, 1000000), 'both')
distalGenes = []
for proxLocus in distalLoci:
distalGenes.append(proxLocus.ID())
overlappingGenes = utils.uniquify(overlappingGenes)
proximalGenes = utils.uniquify(proximalGenes)
distalGenes = utils.uniquify(distalGenes)
allEnhancerGenes = overlappingGenes + proximalGenes + distalGenes
# these checks make sure each gene list is unique.
# technically it is possible for a gene to be overlapping, but not proximal since the
# gene could be longer than the 50kb window, but we'll let that slide
# here
for refID in overlappingGenes:
if proximalGenes.count(refID) == 1:
proximalGenes.remove(refID)
for refID in proximalGenes:
if distalGenes.count(refID) == 1:
distalGenes.remove(refID)
# Now find the closest gene
if len(allEnhancerGenes) == 0:
closestGene = ''
else:
# get enhancerCenter
enhancerCenter = (int(line[2]) + int(line[3])) / 2
# get absolute distance to enhancer center
distList = [abs(enhancerCenter - startDict[geneID]['start'][0])
for geneID in allEnhancerGenes]
# get the ID and convert to name
closestGene = startDict[
allEnhancerGenes[distList.index(min(distList))]]['name']
# NOW WRITE THE ROW FOR THE ENHANCER TABLE
if noFormatTable:
newEnhancerLine = list(line)
newEnhancerLine.append(
join(utils.uniquify([startDict[x]['name'] for x in overlappingGenes]), ','))
newEnhancerLine.append(
join(utils.uniquify([startDict[x]['name'] for x in proximalGenes]), ','))
newEnhancerLine.append(closestGene)
else:
newEnhancerLine = line[0:9]
newEnhancerLine.append(
join(utils.uniquify([startDict[x]['name'] for x in overlappingGenes]), ','))
newEnhancerLine.append(
join(utils.uniquify([startDict[x]['name'] for x in proximalGenes]), ','))
newEnhancerLine.append(closestGene)
newEnhancerLine += line[-2:]
enhancerToGeneTable.append(newEnhancerLine)
# Now grab all overlapping and proximal genes for the gene ordered
# table
overallGeneList += overlappingGenes
for refID in overlappingGenes:
geneDict['overlapping'][refID].append(enhancerString)
rankDict[refID].append(int(line[-2]))
superDict[refID].append(int(line[-1]))
overallGeneList += proximalGenes
for refID in proximalGenes:
geneDict['proximal'][refID].append(enhancerString)
rankDict[refID].append(int(line[-2]))
superDict[refID].append(int(line[-1]))
# End loop through
# Make table by gene
print('MAKING ENHANCER ASSOCIATED GENE TSS COLLECTION')
overallGeneList = utils.uniquify(overallGeneList)
#get the chromLists from the various bams here
cmd = 'samtools idxstats %s' % (rankByBamFile)
idxStats = subprocess.Popen(cmd,stdout=subprocess.PIPE,shell=True)
idxStats= idxStats.communicate()
bamChromList = [line.split('\t')[0] for line in idxStats[0].split('\n')[0:-2]]
if len(controlBamFile) > 0:
cmd = 'samtools idxstats %s' % (controlBamFile)
idxStats = subprocess.Popen(cmd,stdout=subprocess.PIPE,shell=True)
idxStats= idxStats.communicate()
bamChromListControl = [line.split('\t')[0] for line in idxStats[0].split('\n')[0:-2]]
bamChromList = [chrom for chrom in bamChromList if bamChromListControl.count(chrom) != 0]
#now make sure no genes have a bad chrom
overallGeneList = [gene for gene in overallGeneList if bamChromList.count(startDict[gene]['chr']) != 0]
#now make an enhancer collection of all transcripts
enhancerGeneCollection = utils.makeTranscriptCollection(
annotFile, 5000, 5000, 500, overallGeneList)
enhancerGeneGFF = utils.locusCollectionToGFF(enhancerGeneCollection)
# dump the gff to file
enhancerFolder = utils.getParentFolder(enhancerFile)
gffRootName = "%s_TSS_ENHANCER_GENES_-5000_+5000" % (genome)
enhancerGeneGFFFile = "%s%s_%s.gff" % (enhancerFolder, enhancerName,gffRootName)
utils.unParseTable(enhancerGeneGFF, enhancerGeneGFFFile, '\t')
# now we need to run bamToGFF
# Try to use the bamliquidatior_path.py script on cluster, otherwise, failover to local (in path), otherwise fail.
bamliquidator_path = 'bamliquidator_batch'
print('MAPPING SIGNAL AT ENHANCER ASSOCIATED GENE TSS')
# map density at genes in the +/- 5kb tss region
# first on the rankBy bam
bamName = rankByBamFile.split('/')[-1]
mappedRankByFolder = "%s%s_%s_%s/" % (enhancerFolder, enhancerName,gffRootName, bamName)
mappedRankByFile = "%s%s_%s_%s/matrix.txt" % (enhancerFolder,enhancerName, gffRootName, bamName)
cmd = bamliquidator_path + ' --sense . -e 200 --match_bamToGFF -r %s -o %s %s' % (enhancerGeneGFFFile, mappedRankByFolder,rankByBamFile)
print("Mapping rankby bam %s" % (rankByBamFile))
print(cmd)
os.system(cmd)
#check for completion
if utils.checkOutput(mappedRankByFile,0.2,5):
print("SUCCESSFULLY MAPPED TO %s FROM BAM: %s" % (enhancerGeneGFFFile, rankByBamFile))
else:
print("ERROR: FAILED TO MAP %s FROM BAM: %s" % (enhancerGeneGFFFile, rankByBamFile))
sys.exit()
# next on the control bam if it exists
if len(controlBamFile) > 0:
controlName = controlBamFile.split('/')[-1]
mappedControlFolder = "%s%s_%s_%s/" % (
enhancerFolder, enhancerName,gffRootName, controlName)
mappedControlFile = "%s%s_%s_%s/matrix.txt" % (
enhancerFolder, enhancerName,gffRootName, controlName)
cmd = bamliquidator_path + ' --sense . -e 200 --match_bamToGFF -r %s -o %s %s' % (enhancerGeneGFFFile, mappedControlFolder,controlBamFile)
print("Mapping control bam %s" % (controlBamFile))
print(cmd)
os.system(cmd)
#check for completion
if utils.checkOutput(mappedControlFile,0.2,5):
print("SUCCESSFULLY MAPPED TO %s FROM BAM: %s" % (enhancerGeneGFFFile, controlBamFile))
else:
print("ERROR: FAILED TO MAP %s FROM BAM: %s" % (enhancerGeneGFFFile, controlBamFile))
sys.exit()
# now get the appropriate output files
if len(controlBamFile) > 0:
print("CHECKING FOR MAPPED OUTPUT AT %s AND %s" %
(mappedRankByFile, mappedControlFile))
if utils.checkOutput(mappedRankByFile, 1, 1) and utils.checkOutput(mappedControlFile, 1, 1):
print('MAKING ENHANCER ASSOCIATED GENE TSS SIGNAL DICTIONARIES')
signalDict = makeSignalDict(mappedRankByFile, mappedControlFile)
else:
print("NO MAPPING OUTPUT DETECTED")
sys.exit()
else:
print("CHECKING FOR MAPPED OUTPUT AT %s" % (mappedRankByFile))
if utils.checkOutput(mappedRankByFile, 1, 30):
print('MAKING ENHANCER ASSOCIATED GENE TSS SIGNAL DICTIONARIES')
signalDict = makeSignalDict(mappedRankByFile)
else:
print("NO MAPPING OUTPUT DETECTED")
sys.exit()
# use enhancer rank to order
rankOrder = utils.order([min(rankDict[x]) for x in overallGeneList])
usedNames = []
# make a new dict to hold TSS signal by max per geneName
geneNameSigDict = defaultdict(list)
print('MAKING GENE TABLE')
for i in rankOrder:
refID = overallGeneList[i]
geneName = startDict[refID]['name']
if usedNames.count(geneName) > 0 and uniqueGenes == True:
continue
else:
usedNames.append(geneName)
proxEnhancers = geneDict['overlapping'][
refID] + geneDict['proximal'][refID]
superStatus = max(superDict[refID])
enhancerRanks = join([str(x) for x in rankDict[refID]], ',')
enhancerSignal = signalDict[refID]
geneNameSigDict[geneName].append(enhancerSignal)
newLine = [geneName, refID, join(
proxEnhancers, ','), enhancerRanks, superStatus, enhancerSignal]
geneToEnhancerTable.append(newLine)
#utils.unParseTable(geneToEnhancerTable,'/grail/projects/newRose/geneMapper/foo.txt','\t')
print('MAKING ENHANCER TO TOP GENE TABLE')
if noFormatTable:
enhancerToTopGeneTable = [
enhancerToGeneTable[0] + ['TOP_GENE', 'TSS_SIGNAL']]
else:
enhancerToTopGeneTable = [enhancerToGeneTable[0][0:12] + [
'TOP_GENE', 'TSS_SIGNAL'] + enhancerToGeneTable[0][-2:]]
for line in enhancerToGeneTable[1:]:
geneList = []
if noFormatTable:
geneList += line[-3].split(',')
geneList += line[-2].split(',')
else:
geneList += line[10].split(',')
geneList += line[11].split(',')
geneList = utils.uniquify([x for x in geneList if len(x) > 0])
if len(geneList) > 0:
try:
sigVector = [max(geneNameSigDict[x]) for x in geneList]
maxIndex = sigVector.index(max(sigVector))
maxGene = geneList[maxIndex]
maxSig = sigVector[maxIndex]
if maxSig == 0.0:
maxGene = 'NONE'
maxSig = 'NONE'
except ValueError:
if len(geneList) == 1:
maxGene = geneList[0]
maxSig = 'NONE'
else:
maxGene = 'NONE'
maxSig = 'NONE'
else:
maxGene = 'NONE'
maxSig = 'NONE'
if noFormatTable:
newLine = line + [maxGene, maxSig]
else:
newLine = line[0:12] + [maxGene, maxSig] + line[-2:]
enhancerToTopGeneTable.append(newLine)
# resort enhancerToGeneTable
if noFormatTable:
return enhancerToGeneTable, enhancerToTopGeneTable, geneToEnhancerTable
else:
enhancerOrder = utils.order([int(line[-2])
for line in enhancerToGeneTable[1:]])
sortedTable = [enhancerToGeneTable[0]]
sortedTopGeneTable = [enhancerToTopGeneTable[0]]
for i in enhancerOrder:
sortedTable.append(enhancerToGeneTable[(i + 1)])
sortedTopGeneTable.append(enhancerToTopGeneTable[(i + 1)])
return sortedTable, sortedTopGeneTable, geneToEnhancerTable
def make_mycn_stats_table(nb_all_chip_dataFile,outFile):
'''
making a table of conserved mycn peaks w/ some additional stats
mycn and h3k27ac signal is avg. background normalized across 4 samples
active tss defined as the union of all H3K27ac occupied promoters in NB
active enhancers defined as the union of all H3K27ac sites outside of promoters
'''
dataDict = pipeline_dfci.loadDataTable(nb_all_chip_dataFile)
print('SETTING UP OUTPUT TABLE')
outTable = [['PEAK_ID','CHROM','START','STOP','LENGTH','ACTIVE_TSS_OVERLAP','ENHANCER_OVERLAP','CPG_ISLAND_OVERLAP','CPG_ISLAND_FRACTION','GC_FREQ','MYCN_RANK','AVG_MYCN_SIGNAL','AVG_H3K27AC_SIGNAL','CANON_EBOX_COUNT','NONCANON_EBOX_COUNT','TOTAL_EBOX_COUNT','CANON_EXP','NON_CANON_EXP','GABPA_COUNT','GABPA_EXP','GATA_COUNT','GATA_EXP']]
dinuc = nmers(2,['A','T','G','C'])
#input files
mycnSignalFile = '%sHG19_NB_MYCN_CONSERVED_-0_+0_NB_ALL_SIGNAL.txt' % (signalFolder)
h3k27acSignalFile = '%sHG19_NB_MYCN_CONSERVED_-500_+500_NB_ALL_SIGNAL.txt' % (signalFolder)
mycnRankFile = '%smeta_rose/NB_MYCN/NB_MYCN_0KB_STITCHED_ENHANCER_REGION_RANK_CONSERVED.txt' % (projectFolder)
activeGeneFile = '%sHG19_NB_H3K27AC_ACTIVE_UNION.txt' % (geneListFolder)
#note, this is the ucsc hg19 cpg islands extended file
#to download and format run ./beds/download_cpg.sh
cpgFile = '%sbeds/hg19_cpg_islands.bed' % (projectFolder)
enhancerFile = '%smeta_rose/NB_H3K27AC/NB_H3K27AC_AllEnhancers.table.txt' % (projectFolder)
print('LOADING MYCN BINDING DATA')
mycnSignalTable = utils.parseTable(mycnSignalFile,'\t')
#making a signal dictionary for MYCN binding
names_list = ['BE2C_MYCN','KELLY_MYCN','NGP_MYCN','SHEP21_0HR_MYCN_NOSPIKE']
background_list = [dataDict[name]['background'] for name in names_list]
header = mycnSignalTable[0]
chip_columns = [header.index(name) for name in names_list]
background_columns = [header.index(background_name) for background_name in background_list]
mycn_sig_dict = {}
#this only works if the first column are unique identifiers
if len(mycnSignalTable) != len(utils.uniquify([line[0] for line in mycnSignalTable])):
print('Error: Column 1 of must contain unique identifiers.' % (mycnSignalFile))
sys.exit()
for line in mycnSignalTable[1:]:
line_sig = []
for i in range(len(names_list)):
line_sig.append(float(line[chip_columns[i]]) - float(line[background_columns[i]]))
mycn_sig_dict[line[0]] = numpy.mean(line_sig)
print('LOADING MYCN RANK DATA')
mycnRankTable = utils.parseTable(mycnRankFile,'\t')
print('LOADING H3K27AC BINDING DATA')
h3k27acSignalTable = utils.parseTable(h3k27acSignalFile,'\t')
#making a signal dictionary for background subtracted H3K27ac binding
names_list = ['BE2C_H3K27AC','KELLY_H3K27AC','NGP_H3K27AC','SHEP21_0HR_H3K27AC_NOSPIKE']
background_list = [dataDict[name]['background'] for name in names_list]
header = h3k27acSignalTable[0]
chip_columns = [header.index(name) for name in names_list]
background_columns = [header.index(background_name) for background_name in background_list]
h3k27ac_sig_dict = {}
#this only works if the first column are unique identifiers
if len(h3k27acSignalTable) != len(utils.uniquify([line[0] for line in h3k27acSignalTable])):
print('Error: Column 1 of must contain unique identifiers.' % (h3k27acSignalFile))
sys.exit()
for line in h3k27acSignalTable[1:]:
line_sig = []
for i in range(len(names_list)):
line_sig.append(float(line[chip_columns[i]]) - float(line[background_columns[i]]))
h3k27ac_sig_dict[line[0]] = numpy.mean(line_sig)
#making the cpg collection
print('LOADING CPGS ISLANDS')
cpgBed = utils.parseTable(cpgFile,'\t')
cpgLoci = []
for line in cpgBed:
cpgLoci.append(utils.Locus(line[0],line[1],line[2],'.',line[-1]))
cpgCollection = utils.LocusCollection(cpgLoci,50)
#next make the tss collection of active promoters
print('LOADING ACTIVE PROMOTERS')
startDict = utils.makeStartDict(annotFile)
activeTable = utils.parseTable(activeGeneFile,'\t')
tss_1kb_loci = []
for line in activeTable:
tss_1kb_loci.append(utils.makeTSSLocus(line[1],startDict,1000,1000))
tss_1kb_collection = utils.LocusCollection(tss_1kb_loci,50)
#enhancer file
print("LOADING ACTIVE ENHANCERS")
enhancerTable = utils.parseTable(enhancerFile,'\t')
print('STARTING WITH THE FOLLOWING NUMBER OF ENHANCERS IN NB')
print(len(enhancerTable) - 6)
enhancerLoci = []
for line in enhancerTable:
if line[0][0] != '#' and line[0][0] != 'R':
try:
lineLocus = utils.Locus(line[1],int(line[2]),int(line[3]),'.',line[0])
enhancerLoci.append(lineLocus)
except IndexError:
print(line)
sys.exit()
enhancerCollection = utils.LocusCollection(enhancerLoci,50)
print('CLASSIFYING MYCN PEAKS')
ticker = 0
for i in range(1,len(mycnSignalTable)):
if ticker%100 == 0:
print(ticker)
ticker +=1
line = mycnSignalTable[i]
mycn_signal = round(mycn_sig_dict[line[0]],4)
h3k27ac_signal = round(h3k27ac_sig_dict[line[0]],4)
peakID = line[0]
locusString = line[1]
chrom = locusString.split('(')[0]
[start,stop] = [int(x) for x in line[1].split(':')[-1].split('-')]
lineLocus = utils.Locus(chrom,start,stop,'.',peakID)
tssOverlap = 0
if tss_1kb_collection.getOverlap(lineLocus,'both'):
tssOverlap = 1
enhancerOverlap = 0
if enhancerCollection.getOverlap(lineLocus,'both') and tssOverlap == 0:
enhancerOverlap = 1
cpgIslandOverlap = 0
if cpgCollection.getOverlap(lineLocus,'both'):
cpgIslandOverlap = 1
#now do fractional cpgOverlap
overlappingCpGLoci = cpgCollection.getOverlap(lineLocus,'both')
overlappingBases = 0
for locus in overlappingCpGLoci:
cpgStart = max(locus.start(),lineLocus.start())
cpgEnd = min(locus.end(),lineLocus.end())
overlappingBases += (cpgEnd-cpgStart)
overlapFraction = round(float(overlappingBases)/lineLocus.len(),2)
#now get the seq
lineSeq = string.upper(utils.fetchSeq(genomeDirectory,chrom,start,stop,True))
gcFreq = round(float(lineSeq.count('GC') + lineSeq.count('CG'))/len(lineSeq),2)
dinuc_dict = {}
for nmer in dinuc:
dinuc_dict[nmer] = float(lineSeq.count('GC'))/len(lineSeq)
mycnRankLine = mycnRankTable[i]
mycnRank = numpy.mean([float(x) for x in mycnRankLine[6:]])
canonMatchList = re.findall('CACGTG',lineSeq)
canon_count = len(canonMatchList)
eboxMatchList = re.findall('CA..TG',lineSeq)
ebox_count = len(eboxMatchList)
non_canon_count = ebox_count-canon_count
#get the expected values
canon_exp = dinuc_dict['CA']*dinuc_dict['CG']*dinuc_dict['TG']*(len(lineSeq) - 5)
canon_exp = round(canon_exp,2)
notCG = 1- dinuc_dict['CG']
non_exp = dinuc_dict['CA']*notCG*dinuc_dict['TG']*(len(lineSeq) - 5)
non_exp = round(non_exp,2)
#for gata and GABPA
gabpaMatchList = re.findall('CGGAAG',lineSeq) + re.findall('CTTCCG',lineSeq)
gabpa_count = len(gabpaMatchList)
gabpa_exp_f = dinuc_dict['CG'] * dinuc_dict['GA'] * dinuc_dict['AG']*(len(lineSeq) - 5)
gabpa_exp_r = dinuc_dict['CT'] * dinuc_dict['TC'] * dinuc_dict['CG']*(len(lineSeq) - 5)
gabpa_exp = round(gabpa_exp_f,2) + round(gabpa_exp_r,2)
gataMatchList = re.findall('GATAA',lineSeq) + re.findall('TTATC',lineSeq)
gata_count = len(gataMatchList)
an_freq = 1 - dinuc_dict['AA'] - dinuc_dict['AT'] - dinuc_dict['AG'] -dinuc_dict['AC']
cn_freq = 1 - dinuc_dict['CA'] - dinuc_dict['CT'] - dinuc_dict['CG'] -dinuc_dict['CC']
gata_exp_f = dinuc_dict['GA'] * dinuc_dict['TA'] * an_freq*(len(lineSeq) - 5)
gata_exp_r = dinuc_dict['TT'] * dinuc_dict['AT'] * cn_freq*(len(lineSeq) - 5)
gata_exp = round(gata_exp_f,2) + round(gata_exp_r,2)
newLine = [peakID,chrom,start,stop,lineLocus.len(),tssOverlap,enhancerOverlap,cpgIslandOverlap,overlapFraction,gcFreq,mycnRank,mycn_signal,h3k27ac_signal,canon_count,non_canon_count,ebox_count,canon_exp,non_exp,gabpa_count,gabpa_exp,gata_count,gata_exp]
outTable.append(newLine)
utils.unParseTable(outTable,outFile,'\t')
return outFile
def makePeakTable(paramDict,
splitGFFPath,
averageTablePath,
startDict,
geneList,
genomeDirectory,
tads_path=''):
'''
makes the final peak table with ebox info
'''
peakTable = [[
'REGION_ID', 'CHROM', 'START', 'STOP', 'LENGTH', 'TSS', 'CPG',
'CPG_FRACTION', 'GC_FREQ', 'SIGNAL', 'CANON_EBOX_COUNT',
'NON_CANON_EBOX_COUNT', 'TOTAL_EBOX_COUNT', 'OVERLAPPING_GENES',
'PROXIMAL_GENES'
]]
print('LOADING PEAK REGIONS')
peakGFF = utils.parseTable(splitGFFPath, '\t')
print('LOADING BINDING DATA')
signalTable = utils.parseTable(averageTablePath, '\t')
print('LOADING CPGS ISLANDS')
cpgBed = utils.parseTable(paramDict['cpgPath'], '\t')
cpgLoci = []
for line in cpgBed:
cpgLoci.append(utils.Locus(line[0], line[1], line[2], '.', line[-1]))
cpgCollection = utils.LocusCollection(cpgLoci, 50)
print("MAKING TSS COLLECTIONS")
if len(geneList) == 0:
geneList = startDict.keys()
tss_1kb_loci = []
tss_50kb_loci = []
for refID in geneList:
tss_1kb_loci.append(utils.makeTSSLocus(refID, startDict, 1000, 1000))
tss_50kb_loci.append(utils.makeTSSLocus(refID, startDict, 50000,
50000))
#make a 1kb flanking and 50kb flanking collection
tss_1kb_collection = utils.LocusCollection(tss_1kb_loci, 50)
tss_50kb_collection = utils.LocusCollection(tss_50kb_loci, 50)
if len(tads_path) > 0:
print('LOADING TADS FROM %s' % (tads_path))
tad_collection = utils.importBoundRegion(tads_path, 'tad')
use_tads = True
#building a tad dict keyed by tad ID w/ genes in that tad provided
tad_dict = defaultdict(list)
for tss_locus in tss_1kb_loci:
overlapping_tads = tad_collection.getOverlap(tss_locus, 'both')
for tad_locus in overlapping_tads:
tad_dict[tad_locus.ID()].append(tss_locus.ID())
else:
use_tads = False
print('CLASSIFYING PEAKS')
ticker = 0
no_tad_count = 0
for i in range(len(peakGFF)):
if ticker % 1000 == 0:
print(ticker)
ticker += 1
#getting the particulars of the region
gffLine = peakGFF[i]
peakID = gffLine[1]
chrom = gffLine[0]
start = int(gffLine[3])
stop = int(gffLine[4])
lineLocus = utils.Locus(chrom, start, stop, '.', peakID)
#getting the mapped signal
signalLine = signalTable[(i + 1)]
signalVector = [float(x) for x in signalLine[2:]]
#setting up the new line
newLine = [peakID, chrom, start, stop, lineLocus.len()]
#get the tss status from the gff itself (we are able to do this nicely from the split gff code earlier
newLine.append(gffLine[7])
#check cpg status
if cpgCollection.getOverlap(lineLocus, 'both'):
newLine.append(1)
else:
newLine.append(0)
#now do fractional cpgOverlap
overlappingCpGLoci = cpgCollection.getOverlap(lineLocus, 'both')
overlappingBases = 0
for locus in overlappingCpGLoci:
cpgStart = max(locus.start(), lineLocus.start())
cpgEnd = min(locus.end(), lineLocus.end())
overlappingBases += (cpgEnd - cpgStart)
overlapFraction = float(overlappingBases) / lineLocus.len()
newLine.append(round(overlapFraction, 2))
#now get the seq
lineSeq = string.upper(
utils.fetchSeq(genomeDirectory, chrom, start, stop, True))
if len(lineSeq) == 0:
print('UH OH')
print(lineSeq)
print(gffLine)
print(i)
print(chrom)
print(start)
print(stop)
sys.exit()
gcFreq = float(lineSeq.count('GC') +
lineSeq.count('CG')) / len(lineSeq)
newLine.append(gcFreq)
#this is where we add the ChIP-Seq signal
newLine += signalVector
eboxMatchList = re.findall('CA..TG', lineSeq)
if len(eboxMatchList) == 0:
newLine += [0] * 3
else:
totalCount = len(eboxMatchList)
canonCount = eboxMatchList.count('CACGTG')
otherCount = totalCount - canonCount
newLine += [canonCount, otherCount, totalCount]
#now find the overlapping and proximal genes
#here each overlapping gene the tss 1kb locus overlaps the peak
if use_tads:
tad_loci = tad_collection.getOverlap(lineLocus, 'both')
tad_id_list = [tad_locus.ID() for tad_locus in tad_loci]
tad_genes = []
for tad_id in tad_id_list:
tad_genes += tad_dict[tad_id]
if len(tad_genes) == 0:
#print('no tad for this region')
#print(gffLine)
no_tad_count += 1
else:
tad_genes = []
if len(tad_genes) > 0:
overlappingGenes = [
startDict[locus.ID()]['name']
for locus in tss_1kb_collection.getOverlap(lineLocus, 'both')
if tad_genes.count(locus.ID()) > 0
]
proximalGenes = [
startDict[locus.ID()]['name']
for locus in tss_50kb_collection.getOverlap(lineLocus, 'both')
if tad_genes.count(locus.ID()) > 0
]
# print('linked peak to tad genes')
# print([startDict[x]['name'] for x in tad_genes])
# print(tad_id_list)
# print(gffLine)
# print(overlappingGenes)
# print(proximalGenes)
else:
overlappingGenes = [
startDict[locus.ID()]['name']
for locus in tss_1kb_collection.getOverlap(lineLocus, 'both')
]
proximalGenes = [
startDict[locus.ID()]['name']
for locus in tss_50kb_collection.getOverlap(lineLocus, 'both')
]
overlappingGenes = utils.uniquify(overlappingGenes)
#here the tss 50kb locus overlaps the peak
#overlap takes priority over proximal
proximalGenes = [
gene for gene in proximalGenes if overlappingGenes.count(gene) == 0
]
proximalGenes = utils.uniquify(proximalGenes)
overlappingString = string.join(overlappingGenes, ',')
proximalString = string.join(proximalGenes, ',')
newLine += [overlappingString, proximalString]
peakTable.append(newLine)
print('Out of %s regions, %s were assigned to at least 1 tad' %
(len(peakTable), no_tad_count))
return peakTable
def findCanidateTFs(annotationFile, enhancerLoci, expressedNM, expressionDictNM,
bamFile, TFlist, refseqToNameDict, projectFolder, projectName, promoter):
'''
Assign each Super-Enhancer to the closest active TSS to its center
Return a dictionary keyed by TF that points to a list of loci
'''
print 'FINDING CANIDATE TFs'
enhancerAssignment = []
TFtoEnhancerDict = defaultdict(list)
startDict = utils.makeStartDict(annotationFile)
tssLoci = []
for gene in expressedNM:
tssLoci.append(utils.makeTSSLocus(gene,startDict,1000,1000))
tssCollection = utils.LocusCollection(tssLoci,50)
# Loop through enhancers
for enhancer in enhancerLoci:
# If the enhancer overlaps a TSS, save it
overlappingLoci = tssCollection.getOverlap(enhancer, 'both')
overlappingGenes =[]
for overlapLocus in overlappingLoci:
overlappingGenes.append(overlapLocus.ID())
# Find all gene TSS within 100 kb
proximalLoci = tssCollection.getOverlap(utils.makeSearchLocus(enhancer,100000,100000),'both')
proximalGenes =[]
for proxLocus in proximalLoci:
proximalGenes.append(proxLocus.ID())
# If no genes are within 100 kb, find the closest active gene
closestGene = ''
if len(overlappingGenes) == 0 and len(proximalGenes) == 0:
distalLoci = tssCollection.getOverlap(utils.makeSearchLocus(enhancer,1000000,1000000),'both')
distalGenes =[]
for distalLocus in distalLoci:
distalGenes.append(distalLocus.ID())
enhancerCenter = (int(enhancer.start()) + int(enhancer.end())) / 2
distList = [abs(enhancerCenter - startDict[geneID]['start'][0])
for geneID in distalGenes]
if distList:
closestGene = distalGenes[distList.index(min(distList))]
overlappingGenes = utils.uniquify(overlappingGenes)
proximalGenes = utils.uniquify(proximalGenes)
for refID in overlappingGenes:
if proximalGenes.count(refID) == 1:
proximalGenes.remove(refID)
# If a TSS overlaps an enhancer, assign them together
if overlappingGenes:
for gene in overlappingGenes:
if gene in TFlist:
TFtoEnhancerDict[gene].append(enhancer)
enhancerAssignment.append([gene, enhancer.chr(), enhancer.start(), enhancer.end(), enhancer.ID()])
# Otherwise, assign the enhancer to the most active gene in 100 kb
elif not overlappingGenes and proximalGenes:
highestGene = ''
highestActivity = 0
for gene in proximalGenes:
if expressionDictNM[gene] > highestActivity:
highestActivity = expressionDictNM[gene]
highestGene = gene
if highestGene in TFlist:
TFtoEnhancerDict[gene].append(enhancer)
enhancerAssignment.append([gene, enhancer.chr(), enhancer.start(), enhancer.end(), enhancer.ID()])
elif not overlappingGenes and not proximalGenes and closestGene:
if closestGene in TFlist:
gene = closestGene
TFtoEnhancerDict[gene].append(enhancer)
enhancerAssignment.append([gene, enhancer.chr(), enhancer.start(), enhancer.end(), enhancer.ID()])
# Add promoter is it's not contained in the super
if promoter:
for gene in TFtoEnhancerDict.keys():
promoter = utils.Locus(startDict[gene]['chr'], int(startDict[gene]['start'][0]) - 2000,
int(startDict[gene]['start'][0]) + 2000, startDict[gene]['sense'])
overlapBool = False
for enhancer in TFtoEnhancerDict[gene]:
if promoter.overlaps(enhancer):
overlapBool = True
if not overlapBool:
TFtoEnhancerDict[gene].append(promoter)
seAssignmentFile = projectFolder + projectName + '_ENHANCER_ASSIGNMENT.txt'
utils.unParseTable(enhancerAssignment, seAssignmentFile, '\t')
return TFtoEnhancerDict
