1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 161 162 163 164 165 166 167 168 169 170 171 172 173 174 175 176 177 178 179 180 181 182 183 184 185 186 187 188 189 190 191 192 193 194 195 196 197 198 199 200 201 202 203 204 205 206 207 208 209 210 211 212 213 214 215 216 217 218 219 220 221 222 223 224 225
|
#include "sjdbPrepare.h"
#include "ErrorWarning.h"
#include "serviceFuns.cpp"
void sjdbPrepare (SjdbClass &sjdbLoci, Parameters &P, uint nGenomeReal, string outDir, Genome &mapGen, char *Gsj) {
char *G=mapGen.G;
uint *sjdbS=new uint [sjdbLoci.chr.size()];
uint *sjdbE=new uint [sjdbLoci.chr.size()];
uint8 *sjdbMotif=new uint8 [sjdbLoci.chr.size()];
uint8 *sjdbShiftLeft=new uint8 [sjdbLoci.chr.size()];
uint8 *sjdbShiftRight=new uint8 [sjdbLoci.chr.size()];
string chrOld="";
uint iChr=0;
for (uint ii=0;ii<sjdbLoci.chr.size();ii++) {
if (chrOld!=sjdbLoci.chr.at(ii)) {//find numeric value of the chr
for (iChr=0;iChr<mapGen.nChrReal;iChr++) {
if (sjdbLoci.chr.at(ii)==mapGen.chrName[iChr]) break;
};
if (iChr>=mapGen.nChrReal) {
ostringstream errOut;
errOut << "EXITING because of FATAL error, the sjdb chromosome " << sjdbLoci.chr.at(ii) << " is not found among the genomic chromosomes\n";
errOut << "SOLUTION: fix your file(s) --sjdbFileChrStartEnd or --sjdbGTFfile, offending junction:" <<sjdbLoci.chr.at(ii)<<"\t"<<sjdbLoci.start.at(ii)<<"\t"<<sjdbLoci.end.at(ii)<<"\n";
exitWithError(errOut.str(),std::cerr, P.inOut->logMain, EXIT_CODE_INPUT_FILES, P);
};
chrOld=sjdbLoci.chr.at(ii);
};
sjdbS[ii] = sjdbLoci.start.at(ii) + mapGen.chrStart[iChr] - 1;//sj names contain 1-based intron loci
sjdbE[ii] = sjdbLoci.end.at(ii) + mapGen.chrStart[iChr] - 1;
//motifs
if ( G[sjdbS[ii]]==2 && G[sjdbS[ii]+1]==3 && G[sjdbE[ii]-1]==0 && G[sjdbE[ii]]==2 ) {//GTAG
sjdbMotif[ii]=1;
} else if ( G[sjdbS[ii]]==1 && G[sjdbS[ii]+1]==3 && G[sjdbE[ii]-1]==0 && G[sjdbE[ii]]==1 ) {//CTAC
sjdbMotif[ii]=2;
} else if ( G[sjdbS[ii]]==2 && G[sjdbS[ii]+1]==1 && G[sjdbE[ii]-1]==0 && G[sjdbE[ii]]==2 ) {//GCAG
sjdbMotif[ii]=3;
} else if ( G[sjdbS[ii]]==1 && G[sjdbS[ii]+1]==3 && G[sjdbE[ii]-1]==2 && G[sjdbE[ii]]==1 ) {//CTGC
sjdbMotif[ii]=4;
} else if ( G[sjdbS[ii]]==0 && G[sjdbS[ii]+1]==3 && G[sjdbE[ii]-1]==0 && G[sjdbE[ii]]==1 ) {//ATAC
sjdbMotif[ii]=5;
} else if ( G[sjdbS[ii]]==2 && G[sjdbS[ii]+1]==3 && G[sjdbE[ii]-1]==0 && G[sjdbE[ii]]==3 ) {//GTAT
sjdbMotif[ii]=6;
} else {
sjdbMotif[ii]=0;
};
//repeat length: go back and forth around jR to find repeat length
uint jjL=0,jjR=0;
while ( jjL <= sjdbS[ii]-1 && G[sjdbS[ii]-1-jjL]==G[sjdbE[ii]-jjL] && G[sjdbS[ii]-1-jjL]<4 && jjL<255) {//go back
jjL++;
};
sjdbShiftLeft[ii]=jjL;
while ( sjdbS[ii]+jjR < nGenomeReal && G[sjdbS[ii]+jjR]==G[sjdbE[ii]+1+jjR] && G[sjdbS[ii]+jjR]<4 && jjR<255) {//go forward
jjR++;
};
sjdbShiftRight[ii]=jjR;
if (jjR==255 || jjL==255) {
P.inOut->logMain << "WARNING: long repeat for junction # " << ii+1 <<" : " \
<< sjdbLoci.chr.at(ii) <<" "<<sjdbS[ii] - mapGen.chrStart[iChr] + 1 <<" "<< sjdbE[ii] - mapGen.chrStart[iChr] + 1 \
<< "; left shift = "<< (int) sjdbShiftLeft[ii] <<"; right shift = "<< (int) sjdbShiftRight[ii] <<"\n";
};
sjdbS[ii]-=sjdbShiftLeft[ii];
sjdbE[ii]-=sjdbShiftLeft[ii];
};
//sort sjdb
uint *sjdbSort=new uint [sjdbLoci.chr.size()*3];
for (uint ii=0;ii<sjdbLoci.chr.size();ii++) {
uint shift1=0;
switch (sjdbLoci.str.at(ii)) {
case '+':
shift1=0;
break;
case '-':
shift1=nGenomeReal;
break;
default:
shift1=2*nGenomeReal;
};
sjdbSort[ii*3]=sjdbS[ii]+shift1; //separate sorting of +/- strand
sjdbSort[ii*3+1]=sjdbE[ii]+shift1;
sjdbSort[ii*3+2]=ii;
};
qsort((void *) sjdbSort, sjdbLoci.chr.size(), sizeof(uint)*3, funCompareUint2);
uint *I=new uint [sjdbLoci.chr.size()];
uint nsj=0;
for (uint ii=0;ii<sjdbLoci.chr.size();ii++) {
uint isj=sjdbSort[ii*3+2];//index of the next sorted junction
uint isj0;
if (nsj>0)
{
isj0=I[nsj-1]; //index of the last recorded junctions
};
if (nsj==0 || sjdbS[isj]!=sjdbS[isj0] || sjdbE[isj]!=sjdbE[isj0])
{//different intron coordinates
I[nsj++]=isj;// add new junction
} else if (sjdbLoci.priority.at(isj)<sjdbLoci.priority.at(isj0))
{//new junction has lower priority
//do nothing, i.e. keep the old junction
} else if (sjdbLoci.priority.at(isj)>sjdbLoci.priority.at(isj0))
{//new junction has higher priority
I[nsj-1]=isj;//replace the old junction
} else if ( (sjdbMotif[isj]>0 && sjdbMotif[isj0]==0) \
|| ( ((sjdbMotif[isj]>0) == (sjdbMotif[isj0]>0)) && sjdbShiftLeft[isj]<sjdbShiftLeft[isj0]) ) {
//new and old junctions have the same priority
//new junction is canonical or left-most, so it wins
I[nsj-1]=isj;//replace the old junction
// else - keep the old junction, do not add the new one
};
};
//sort again, after returning canonical junctions back to original loci:
for (uint ii=0;ii<nsj;ii++) {
sjdbSort[ii*3] =sjdbS[I[ii]] + (sjdbMotif[I[ii]]==0 ? 0 : sjdbShiftLeft[I[ii]]);
sjdbSort[ii*3+1]=sjdbE[I[ii]] + (sjdbMotif[I[ii]]==0 ? 0 : sjdbShiftLeft[I[ii]]);
sjdbSort[ii*3+2]=I[ii];
};
qsort((void *) sjdbSort, nsj, sizeof(uint)*3, funCompareUint2);
mapGen.sjdbStart=new uint [nsj];
mapGen.sjdbEnd=new uint [nsj];
mapGen.sjdbMotif=new uint8 [nsj];
mapGen.sjdbShiftLeft=new uint8 [nsj];
mapGen.sjdbShiftRight=new uint8 [nsj];
mapGen.sjdbStrand=new uint8 [nsj];
uint nsj1=0;
for (uint ii=0;ii<nsj;ii++) {
uint isj=sjdbSort[ii*3+2];
if ( nsj1>0 && mapGen.sjdbStart[nsj1-1]==sjdbSort[ii*3] && mapGen.sjdbEnd[nsj1-1]==sjdbSort[ii*3+1] ) {//same loci on opposite strands
uint isj0=sjdbSort[(ii-1)*3+2];
if (sjdbLoci.priority.at(isj)<sjdbLoci.priority.at(isj0))
{//new junction has lower priority
continue;//keep old junction, do not add new
} else if (sjdbLoci.priority.at(isj)>sjdbLoci.priority.at(isj0))
{//new junction has higher priority
nsj1--;//replace the old junction with the new one
} else if (mapGen.sjdbStrand[nsj1-1]>0 && sjdbLoci.str.at(isj)=='.')
{//new junction strand is not defined
continue;
} else if (mapGen.sjdbStrand[nsj1-1]==0 && sjdbLoci.str.at(isj)!='.')
{//old junction strand is not defined
nsj1--; //replace old with new
} else if (mapGen.sjdbMotif[nsj1-1]==0 && sjdbMotif[isj]==0)
{//both are non-canonical (on opposite strand)
mapGen.sjdbStrand[nsj1-1]=0;//do not record new junction, keep old with undefined strand
continue;
} else if ( (mapGen.sjdbMotif[nsj1-1]>0 && sjdbMotif[isj]==0) ||(mapGen.sjdbMotif[nsj1-1]%2 == (2-mapGen.sjdbStrand[nsj1-1])) ){//both strands defined, both junctions canonical
//old junction is canonical, new is not, OR old junction is on correct strand
continue;
} else {
//new junction is on correct strand, replace the old one
nsj1--;
};
};
//record junction
mapGen.sjdbStart[nsj1]=sjdbSort[ii*3];
mapGen.sjdbEnd[nsj1]=sjdbSort[ii*3+1];
mapGen.sjdbMotif[nsj1]=sjdbMotif[isj];
mapGen.sjdbShiftLeft[nsj1]=sjdbShiftLeft[isj];
mapGen.sjdbShiftRight[nsj1]=sjdbShiftRight[isj];
if (sjdbLoci.str.at(isj)=='+') {
mapGen.sjdbStrand[nsj1]=1;
} else if (sjdbLoci.str.at(isj)=='-') {
mapGen.sjdbStrand[nsj1]=2;
} else {
if (mapGen.sjdbMotif[nsj1]==0) {//strand un-defined
mapGen.sjdbStrand[nsj1]=0;
} else {
mapGen.sjdbStrand[nsj1]=2-mapGen.sjdbMotif[nsj1]%2;
};
};
nsj1++;
};
mapGen.sjdbN=nsj1;
mapGen.sjDstart = new uint [mapGen.sjdbN];
mapGen.sjAstart = new uint [mapGen.sjdbN];
ofstream sjdbInfo((outDir+"/sjdbInfo.txt").c_str());
ofstream sjdbList ((outDir+"/sjdbList.out.tab").c_str());
char strandChar[3]={'.','+','-'};
//first line is some general useful information
sjdbInfo << mapGen.sjdbN <<"\t"<< mapGen.sjdbOverhang <<"\n";
uint sjGstart=0;
for (uint ii=0;ii<mapGen.sjdbN;ii++)
{//add sjdb sequence to genome
mapGen.sjDstart[ii] = mapGen.sjdbStart[ii] - mapGen.sjdbOverhang;
mapGen.sjAstart[ii] = mapGen.sjdbEnd[ii] + 1;
if (mapGen.sjdbMotif[ii]==0) {//shift non-canonical junctions back to their true coordinates
mapGen.sjDstart[ii] += mapGen.sjdbShiftLeft[ii];
mapGen.sjAstart[ii] += mapGen.sjdbShiftLeft[ii];
};
memcpy(Gsj+sjGstart,G+mapGen.sjDstart[ii],mapGen.sjdbOverhang);//sjdbStart contains 1-based intron loci
memcpy(Gsj+sjGstart+mapGen.sjdbOverhang,G+mapGen.sjAstart[ii],mapGen.sjdbOverhang);//sjdbStart contains 1-based intron loci
sjGstart += mapGen.sjdbLength;
Gsj[sjGstart-1]=GENOME_spacingChar;//spacing char between the sjdb seqs
sjdbInfo << mapGen.sjdbStart[ii] <<"\t"<< mapGen.sjdbEnd[ii] <<"\t"<<(int) mapGen.sjdbMotif[ii] <<"\t"<<(int) mapGen.sjdbShiftLeft[ii] <<"\t"<<(int) mapGen.sjdbShiftRight[ii]<<"\t"<<(int) mapGen.sjdbStrand[ii] <<"\n";
uint chr1=mapGen.chrBin[ mapGen.sjdbStart[ii] >> P.pGe.gChrBinNbits];
sjdbList << mapGen.chrName[chr1]<< "\t" << mapGen.sjdbStart[ii]-mapGen.chrStart[chr1] + 1 + (mapGen.sjdbMotif[ii]>0 ? 0:mapGen.sjdbShiftLeft[ii]) \
<< "\t"<< mapGen.sjdbEnd[ii]-mapGen.chrStart[chr1] + 1 + (mapGen.sjdbMotif[ii]>0 ? 0:mapGen.sjdbShiftLeft[ii]) \
<< "\t"<< strandChar[mapGen.sjdbStrand[ii]]<<"\n";
};
sjdbInfo.close();
sjdbList.close();
};
|