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1 package edu.unc.genomics.nucleosomes;
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2
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3 import java.io.BufferedWriter;
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4 import java.io.IOException;
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5 import java.nio.charset.Charset;
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6 import java.nio.file.Files;
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7 import java.nio.file.Path;
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8 import java.util.Iterator;
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9
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10 import org.apache.log4j.Logger;
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11 import org.broad.igv.bbfile.WigItem;
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12
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13 import com.beust.jcommander.Parameter;
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14 import edu.unc.genomics.CommandLineTool;
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15 import edu.unc.genomics.CommandLineToolException;
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16 import edu.unc.genomics.ReadablePathValidator;
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17 import edu.unc.genomics.io.WigFile;
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18 import edu.unc.genomics.io.WigFileException;
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19 import edu.unc.utils.SortUtils;
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20
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21 public class GreedyCaller extends CommandLineTool {
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22
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23 private static final Logger log = Logger.getLogger(GreedyCaller.class);
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24
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25 private static final int CHUNK_SIZE = 500_000;
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26
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27 @Parameter(names = {"-d", "--dyads"}, description = "Dyad counts file", required = true, validateWith = ReadablePathValidator.class)
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28 public WigFile dyadsFile;
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29 @Parameter(names = {"-s", "--smoothed"}, description = "Smoothed dyad counts file", required = true, validateWith = ReadablePathValidator.class)
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30 public WigFile smoothedDyadsFile;
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31 @Parameter(names = {"-n", "--size"}, description = "Nucleosome size (bp)")
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32 public int nucleosomeSize = 147;
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33 @Parameter(names = {"-o", "--output"}, description = "Output file", required = true)
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34 public Path outputFile;
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35
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36 public void run() throws IOException {
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37 int halfNuc = nucleosomeSize / 2;
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38
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39 try (BufferedWriter writer = Files.newBufferedWriter(outputFile, Charset.defaultCharset())) {
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40 for (String chr : smoothedDyadsFile.chromosomes()) {
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41 log.debug("Processing chromosome "+chr);
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42 int chunkStart = smoothedDyadsFile.getChrStart(chr);
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43 int chrStop = smoothedDyadsFile.getChrStop(chr);
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44 while (chunkStart < chrStop) {
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45 int chunkStop = chunkStart + CHUNK_SIZE;
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46 int paddedStart = chunkStart - nucleosomeSize;
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47 int paddedStop = chunkStop + nucleosomeSize;
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48 log.debug("Processing chunk "+chunkStart+"-"+chunkStop);
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49
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50 log.debug("Loading data and sorting");
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51 Iterator<WigItem> dyadsIter;
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52 Iterator<WigItem> smoothedIter;
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53 try {
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54 dyadsIter = dyadsFile.query(chr, paddedStart, paddedStop);
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55 smoothedIter = smoothedDyadsFile.query(chr, paddedStart, paddedStop);
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56 } catch (IOException | WigFileException e) {
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57 e.printStackTrace();
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58 throw new CommandLineToolException("Error loading data from Wig file");
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59 }
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60
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61 float[] dyads = WigFile.flattenData(dyadsIter, paddedStart, paddedStop);
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62 float[] smoothed = WigFile.flattenData(smoothedIter, paddedStart, paddedStop);
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63 int[] sortedIndices = SortUtils.indexSort(smoothed);
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64
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65 // Proceed through the data in descending order
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66 log.debug("Calling nucleosomes");
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67 for (int j = sortedIndices.length; j >= 0; j++) {
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68 int i = sortedIndices[j];
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69 int dyad = paddedStart + i;
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70
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71 if (smoothed[i] > 0) {
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72 int nucStart = Math.max(1, dyad-halfNuc);
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73 int nucStop = Math.min(dyad+halfNuc, chrStop);
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74 NucleosomeCall call = new NucleosomeCall(chr, nucStart, nucStop);
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75 call.setDyad(dyad);
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76
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77 double occupancy = 0;
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78 double weightedSum = 0;
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79 double smoothedSum = 0;
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80 double sumOfSquares = 0;
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81 for (int bp = nucStart; bp <= nucStop; bp++) {
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82 occupancy += dyads[bp-paddedStart];
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83 weightedSum += bp * dyads[bp-paddedStart];
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84 smoothedSum += smoothed[bp-paddedStart];
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85 sumOfSquares += bp * bp * dyads[bp-paddedStart];
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86 }
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87 call.setOccupancy(occupancy);
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88
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89 if (occupancy > 0) {
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90 call.setDyadMean(Math.round(weightedSum/occupancy));
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91 call.setConditionalPosition(smoothed[i] / smoothedSum);
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92 double variance = (sumOfSquares - weightedSum*call.getDyadMean()) / occupancy;
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93 call.setDyadStdev(Math.sqrt(variance));
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94
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95 // Only write nucleosomes within the current chunk to disk
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96 if (chunkStart <= dyad && dyad <= chunkStop) {
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97 writer.write(call.toString());
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98 writer.newLine();
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99 }
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100
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101 // Don't allow nucleosome calls overlapping this nucleosome
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102 int low = Math.max(i-nucleosomeSize, 0);
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103 int high = Math.min(i-nucleosomeSize, paddedStop-1);
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104 for (int k = low; k <= high; k++) {
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105 smoothed[k] = 0;
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106 }
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107 }
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108 }
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109 }
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110
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111 chunkStart = chunkStop + 1;
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112 }
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113 }
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114 }
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115 }
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116
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117 public static void main(String[] args) {
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118 new GreedyCaller().instanceMain(args);
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119 }
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120 } |