1 | #!/usr/bin/perl -w |
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2 | |
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3 | # a program to compute the frequency of each motif at each window in both upstream and downstream sequences flanking indels |
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4 | # in a chromosome/genome. |
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5 | # the first input is a TABULAR format file containing the motif names and sequences, such that the file consists of two |
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6 | # columns: the left column represents the motif names and the right column represents the motif sequence, one line per motif. |
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7 | # the second input is a TABULAR format file containing the upstream and downstream sequences flanking indels, one line per indel. |
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8 | # the fourth input is an integer number representing the window size according to which the upstream and downstream sequences |
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9 | # flanking each indel will be divided. |
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10 | # the first output is a TABULAR format file containing the windows into which both upstream and downstream sequences flanking |
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11 | # indels are divided. |
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12 | # the second output is a TABULAR format file containing the motifs and their corresponding frequencies at each window in both |
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13 | # upstream and downstream sequences flanking indels, one line per motif. |
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14 | |
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15 | use strict; |
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16 | use warnings; |
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17 | |
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18 | #variable to handle the falnking sequences information |
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19 | my $sequence = ""; |
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20 | my $upstreamFlankingSequence = ""; |
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21 | my $downstreamFlankingSequence = ""; |
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22 | my $discardedSequenceLength = 0; |
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23 | my $lengthOfDownstreamFlankingSequenceAfterTrimming = 0; |
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24 | |
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25 | #variable to handle the window information |
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26 | my $window = ""; |
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27 | my $windowStartIndex = 0; |
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28 | my $windowNumber = 0; |
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29 | my $totalWindowsNumber = 0; |
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30 | my $totalNumberOfWindowsInUpstreamSequence = 0; |
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31 | my $totalNumberOfWindowsInDownstreamSequence = 0; |
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32 | my $totalWindowsNumberInBothFlankingSequences = 0; |
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33 | my $totalWindowsNumberInMotifCountersTwoDimArray = 0; |
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34 | my $upstreamAndDownstreamFlankingSequencesWindows = ""; |
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35 | |
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36 | #variable to handle the motif information |
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37 | my $motif = ""; |
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38 | my $motifSequence = ""; |
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39 | my $motifNumber = 0; |
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40 | my $totalMotifsNumber = 0; |
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41 | |
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42 | #arrays to sotre window and motif data |
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43 | my @windowsArray = (); |
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44 | my @motifNamesArray = (); |
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45 | my @motifSequencesArray = (); |
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46 | my @motifCountersTwoDimArray = (); |
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47 | |
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48 | #variables to store line counter values |
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49 | my $lineCounter1 = 0; |
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50 | my $lineCounter2 = 0; |
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51 | |
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52 | # check to make sure having correct files |
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53 | my $usage = "usage: compute_motifs_frequency.pl [TABULAR.in] [TABULAR.in] [windowSize] [TABULAR.out] [TABULAR.out]\n"; |
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54 | die $usage unless @ARGV == 5; |
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55 | |
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56 | #get the input and output arguments |
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57 | my $motifsInputFile = $ARGV[0]; |
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58 | my $indelFlankingSequencesInputFile = $ARGV[1]; |
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59 | my $windowSize = $ARGV[2]; |
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60 | my $indelFlankingSequencesWindowsOutputFile = $ARGV[3]; |
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61 | my $motifFrequenciesOutputFile = $ARGV[4]; |
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62 | |
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63 | #open the input and output files |
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64 | open (INPUT1, "<", $motifsInputFile) || die("Could not open file $motifsInputFile \n"); |
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65 | open (INPUT2, "<", $indelFlankingSequencesInputFile) || die("Could not open file $indelFlankingSequencesInputFile \n"); |
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66 | open (OUTPUT1, ">", $indelFlankingSequencesWindowsOutputFile) || die("Could not open file $indelFlankingSequencesWindowsOutputFile \n"); |
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67 | open (OUTPUT2, ">", $motifFrequenciesOutputFile) || die("Could not open file $motifFrequenciesOutputFile \n"); |
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68 | |
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69 | #store the motifs input file in the array @motifsData |
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70 | my @motifsData = <INPUT1>; |
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71 | |
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72 | #iterated through the motifs (lines) of the motifs input file |
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73 | foreach $motif (@motifsData){ |
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74 | chomp ($motif); |
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75 | #print ($motif . "\n"); |
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76 | |
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77 | #split the motif data into its name and its sequence |
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78 | my @motifNameAndSequenceArray = split(/\t/, $motif); |
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79 | |
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80 | #store the name of the motif into the array @motifNamesArray |
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81 | push @motifNamesArray, $motifNameAndSequenceArray[0]; |
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82 | |
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83 | #store the sequence of the motif into the array @motifSequencesArray |
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84 | push @motifSequencesArray, $motifNameAndSequenceArray[1]; |
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85 | } |
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86 | |
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87 | #compute the size of the motif names array |
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88 | $totalMotifsNumber = @motifNamesArray; |
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89 | |
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90 | #store the input file in the array @sequencesData |
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91 | my @sequencesData = <INPUT2>; |
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92 | |
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93 | #iterated through the sequences of the second input file in order to create windwos file |
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94 | foreach $sequence (@sequencesData){ |
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95 | chomp ($sequence); |
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96 | $lineCounter1++; |
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97 | |
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98 | my @indelAndSequenceArray = split(/\t/, $sequence); |
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99 | |
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100 | #get the upstream falnking sequence |
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101 | $upstreamFlankingSequence = $indelAndSequenceArray[3]; |
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102 | |
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103 | #if the window size is 0, then the whole upstream will be one window only |
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104 | if ($windowSize == 0){ |
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105 | $totalNumberOfWindowsInUpstreamSequence = 1; |
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106 | $windowSize = length ($upstreamFlankingSequence); |
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107 | } |
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108 | else{ |
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109 | #compute the total number of windows into which the upstream flanking sequence will be divided |
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110 | $totalNumberOfWindowsInUpstreamSequence = length ($upstreamFlankingSequence) / $windowSize; |
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111 | |
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112 | #compute the length of the subsequence to be discared from the upstream flanking sequence if any |
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113 | $discardedSequenceLength = length ($upstreamFlankingSequence) % $windowSize; |
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114 | |
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115 | #check if the sequence could be split into windows of equal sizes |
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116 | if ($discardedSequenceLength != 0) { |
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117 | #trim the upstream flanking sequence |
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118 | $upstreamFlankingSequence = substr($upstreamFlankingSequence, $discardedSequenceLength); |
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119 | } |
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120 | } |
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121 | |
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122 | #split the upstream flanking sequence into windows |
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123 | for ($windowNumber = 0; $windowNumber < $totalNumberOfWindowsInUpstreamSequence; $windowNumber++){ |
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124 | $windowStartIndex = $windowNumber * $windowSize; |
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125 | print OUTPUT1 (substr($upstreamFlankingSequence, $windowStartIndex, $windowSize) . "\t"); |
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126 | } |
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127 | |
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128 | #add a column representing the indel |
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129 | print OUTPUT1 ("indel" . "\t"); |
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130 | |
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131 | #get the downstream falnking sequence |
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132 | $downstreamFlankingSequence = $indelAndSequenceArray[4]; |
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133 | |
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134 | #if the window size is 0, then the whole upstream will be one window only |
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135 | if ($windowSize == 0){ |
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136 | $totalNumberOfWindowsInDownstreamSequence = 1; |
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137 | $windowSize = length ($downstreamFlankingSequence); |
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138 | } |
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139 | else{ |
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140 | #compute the total number of windows into which the downstream flanking sequence will be divided |
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141 | $totalNumberOfWindowsInDownstreamSequence = length ($downstreamFlankingSequence) / $windowSize; |
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142 | |
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143 | #compute the length of the subsequence to be discared from the upstream flanking sequence if any |
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144 | $discardedSequenceLength = length ($downstreamFlankingSequence) % $windowSize; |
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145 | |
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146 | #check if the sequence could be split into windows of equal sizes |
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147 | if ($discardedSequenceLength != 0) { |
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148 | #compute the length of the sequence to be discarded |
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149 | $lengthOfDownstreamFlankingSequenceAfterTrimming = length ($downstreamFlankingSequence) - $discardedSequenceLength; |
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150 | |
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151 | #trim the downstream flanking sequence |
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152 | $downstreamFlankingSequence = substr($downstreamFlankingSequence, 0, $lengthOfDownstreamFlankingSequenceAfterTrimming); |
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153 | } |
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154 | } |
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155 | |
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156 | #split the downstream flanking sequence into windows |
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157 | for ($windowNumber = 0; $windowNumber < $totalNumberOfWindowsInDownstreamSequence; $windowNumber++){ |
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158 | $windowStartIndex = $windowNumber * $windowSize; |
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159 | print OUTPUT1 (substr($downstreamFlankingSequence, $windowStartIndex, $windowSize) . "\t"); |
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160 | } |
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161 | |
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162 | print OUTPUT1 ("\n"); |
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163 | } |
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164 | |
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165 | #compute the total number of windows on both upstream and downstream sequences flanking the indel |
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166 | $totalWindowsNumberInBothFlankingSequences = $totalNumberOfWindowsInUpstreamSequence + $totalNumberOfWindowsInDownstreamSequence; |
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167 | |
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168 | #add an additional cell to store the name of the motif and another one for the indel itself |
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169 | $totalWindowsNumberInMotifCountersTwoDimArray = $totalWindowsNumberInBothFlankingSequences + 1 + 1; |
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170 | |
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171 | #initialize the two dimensional array $motifCountersTwoDimArray. the first column will be initialized with motif names |
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172 | for ($motifNumber = 0; $motifNumber < $totalMotifsNumber; $motifNumber++){ |
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173 | |
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174 | for ($windowNumber = 0; $windowNumber < $totalWindowsNumberInMotifCountersTwoDimArray; $windowNumber++){ |
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175 | |
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176 | if ($windowNumber == 0){ |
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177 | $motifCountersTwoDimArray [$motifNumber] [0] = $motifNamesArray[$motifNumber]; |
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178 | } |
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179 | elsif ($windowNumber == $totalNumberOfWindowsInUpstreamSequence + 1){ |
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180 | $motifCountersTwoDimArray [$motifNumber] [$windowNumber] = "indel"; |
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181 | } |
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182 | else{ |
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183 | $motifCountersTwoDimArray [$motifNumber] [$windowNumber] = 0; |
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184 | } |
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185 | } |
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186 | } |
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187 | |
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188 | close(OUTPUT1); |
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189 | |
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190 | #open the file the contains the windows of the upstream and downstream flanking sequences, which is actually the first output file |
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191 | open (INPUT3, "<", $indelFlankingSequencesWindowsOutputFile) || die("Could not open file $indelFlankingSequencesWindowsOutputFile \n"); |
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192 | |
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193 | #store the first output file containing the windows of both upstream and downstream flanking sequences in the array @windowsData |
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194 | my @windowsData = <INPUT3>; |
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195 | |
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196 | #iterated through the lines of the first output file. Each line represents |
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197 | #the windows of the upstream and downstream flanking sequences of an indel |
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198 | foreach $upstreamAndDownstreamFlankingSequencesWindows (@windowsData){ |
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199 | |
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200 | chomp ($upstreamAndDownstreamFlankingSequencesWindows); |
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201 | $lineCounter2++; |
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202 | |
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203 | #split both upstream and downstream flanking sequences into their windows |
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204 | my @windowsArray = split(/\t/, $upstreamAndDownstreamFlankingSequencesWindows); |
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205 | |
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206 | $totalWindowsNumber = @windowsArray; |
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207 | |
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208 | #iterate through the windows to search for matched motifs and increment their corresponding counters accordingly |
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209 | WINDOWS: |
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210 | for ($windowNumber = 0; $windowNumber < $totalWindowsNumber; $windowNumber++){ |
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211 | |
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212 | #get the window |
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213 | $window = $windowsArray[$windowNumber]; |
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214 | |
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215 | #if the window is the one that contains the indel, then skip the indel window |
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216 | if ($window eq "indel") { |
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217 | next WINDOWS; |
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218 | } |
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219 | else{ #iterated through the motif sequences to check their occurrences in the winodw |
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220 | #and increment their corresponding counters accordingly |
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221 | |
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222 | for ($motifNumber = 0; $motifNumber < $totalMotifsNumber; $motifNumber++){ |
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223 | #get the motif sequence |
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224 | $motifSequence = $motifSequencesArray[$motifNumber]; |
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225 | |
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226 | #if the motif is found in the window, then increment its corresponding counter |
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227 | if ($window =~ m/$motifSequence/i){ |
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228 | $motifCountersTwoDimArray [$motifNumber] [$windowNumber + 1]++; |
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229 | } |
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230 | } |
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231 | } |
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232 | } |
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233 | } |
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234 | |
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235 | #store the motif counters values in the second output file |
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236 | for ($motifNumber = 0; $motifNumber < $totalMotifsNumber; $motifNumber++){ |
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237 | |
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238 | for ($windowNumber = 0; $windowNumber <= $totalWindowsNumber; $windowNumber++){ |
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239 | |
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240 | print OUTPUT2 $motifCountersTwoDimArray [$motifNumber] [$windowNumber] . "\t"; |
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241 | #print ($motifCountersTwoDimArray [$motifNumber] [$windowNumber] . " "); |
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242 | } |
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243 | print OUTPUT2 "\n"; |
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244 | #print ("\n"); |
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245 | } |
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246 | |
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247 | #close the input and output files |
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248 | close(OUTPUT2); |
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249 | close(OUTPUT1); |
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250 | close(INPUT3); |
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251 | close(INPUT2); |
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252 | close(INPUT1); |
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