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1 package ppp;
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2
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3 use strict;
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4 use warnings;
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5 use FindBin;
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6 use lib $FindBin::Bin;
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7 use Rcall qw ( histogram );
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8 use Math::CDF;
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9
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10 use Exporter;
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11 our @ISA = qw( Exporter );
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12 our @EXPORT_OK = qw( &ping_pong_partners );
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13
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14 sub ping_pong_partners
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15 {
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16 my ( $TE_fai, $sam, $dir, $max ) = @_;
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17 my ( $hashRef, $dupRef, $hasPpp ) = count_mapped ( $TE_fai, $sam );
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18 my ( %num_per_overlap_size, $overlap_number, $reverseR, $begRev, $endRev, $sensR, $begSens, $endSens, $snum, $rnum, $overlap );
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19 my ( $SP, $AP, $SN, $AN, $txt );
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20 my $flag = 0;
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21 my @distri_overlap = (); my @overlaps_names = ();
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22
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23 open my $ppp_f, '>', $dir."ppp.txt" || die "cannot create ppp.txt $!\n";
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24 foreach my $k ( sort keys %{$hashRef} )
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25 {
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26 my $v = $hashRef->{$k};
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27 my $TE_dir = $dir.$k.'/';
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28
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29 %num_per_overlap_size = (); $overlap_number = 0;
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30 $flag = 0;
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31 for ( my $i = 0; $i <= $#{$v->[1]} ; $i++ )
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32 {
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33 $reverseR = ${$v->[1]}[$i] ;
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34 $begRev = $reverseR->[0];
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35 $endRev = $begRev + length($reverseR->[1]) - 1;
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36
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37 my $revR = reverse($reverseR->[1]);
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38 $revR =~ tr/atgcuATGCU/tacgaTACGA/;
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39
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40 for ( my $j = 0; $j <= $#{$v->[0]}; $j++ )
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41 {
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42 $sensR = ${$v->[0]}[$j];
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43 $begSens = $sensR->[0];
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44 $endSens = $begSens + length($sensR->[1]) - 1;
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45
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46 if ( $begSens <= $endRev && $endSens > $endRev )
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47 {
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48 $flag = 1;
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49 mkdir $TE_dir;
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50 open $txt, '>', $TE_dir.'overlap_size.txt' || die "cannot open repartition\n";
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51
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52 $overlap = $endRev - $begSens + 1;
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53 $snum = $dupRef->{$sensR->[0].$sensR->[1].$sensR->[2].$sensR->[3]};
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54 $rnum = $dupRef->{$reverseR->[0].$reverseR->[1].$reverseR->[2].$reverseR->[3]};
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55
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56 if ( $overlap == 10 )
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57 {
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58 $hasPpp->{ $sensR->[0].$sensR->[1].$sensR->[2].$sensR->[3] } = 1;
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59 $hasPpp->{ $reverseR->[0].$reverseR->[1].$reverseR->[2].$reverseR->[3] } = 1;
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60 }
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61 next if $overlap > $max;
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62 if ( $snum < $rnum )
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63 {
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64 $num_per_overlap_size{$overlap} += $snum;
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65 $overlap_number += $snum;
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66 }
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67 else
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68 {
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69 $num_per_overlap_size{$overlap} += $rnum ;
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70 $overlap_number += $rnum ;
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71 }
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72 }
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73 }
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74 }
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75 if ( $max != 0 )
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76 {
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77 my @overlaps = ();
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78 push @overlaps_names, $k;
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79 for my $i (1..$max)
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80 {
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81 $num_per_overlap_size{$i} = 0 unless exists( $num_per_overlap_size{$i} );
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82 push @overlaps, $num_per_overlap_size{$i};
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83 }
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84 push @distri_overlap, \@overlaps;
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85 }
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86
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87 if ( $flag == 1 )
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88 {
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89 open $AP, '>', $TE_dir."antisensPPP.txt" || die "cannot create antisensPPP\n";
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90 open $AN, '>', $TE_dir."antisens.txt" || die "cannot create antisens\n";
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91 for ( my $i = 0; $i <= $#{$v->[1]} ; $i++ )
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92 {
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93 $reverseR = ${$v->[1]}[$i] ;
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94 my $revR = reverse($reverseR->[1]);
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95 $revR =~ tr/atgcuATGCU/tacgaTACGA/;
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96 $rnum = $dupRef->{$reverseR->[0].$reverseR->[1].$reverseR->[2].$reverseR->[3]};
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97 if ( $hasPpp->{ $reverseR->[0].$reverseR->[1].$reverseR->[2].$reverseR->[3] } == 1 )
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98 {
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99 print $AP ">$reverseR->[0]|$reverseR->[2]|$reverseR->[3]|$rnum\n$revR\n";
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100 }
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101 else
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102 {
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103 print $AN ">$reverseR->[0]|$reverseR->[2]|$reverseR->[3]|$rnum\n$revR\n";
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104 }
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105 }
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106 close $AP; close $AN;
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107
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108 open $SP, '>', $TE_dir."sensPPP.txt" || die "cannot create sensPPP\n";
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109 open $SN, '>', $TE_dir."sens.txt" || die "cannot create sens\n";
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110 for ( my $j = 0; $j <= $#{$v->[0]}; $j++ )
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111 {
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112 $sensR = ${$v->[0]}[$j];
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113 $snum = $dupRef->{$sensR->[0].$sensR->[1].$sensR->[2].$sensR->[3]};
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114 if ( $hasPpp->{ $sensR->[0].$sensR->[1].$sensR->[2].$sensR->[3] } == 1 )
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115 {
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116 print $SP ">$sensR->[0]|$sensR->[2]|$sensR->[3]|$snum\n$sensR->[1]\n";
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117 }
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118 else
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119 {
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120 print $SN ">$sensR->[0]|$sensR->[2]|$sensR->[3]|$snum\n$sensR->[1]\n";
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121 }
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122 }
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123 close $SP; close $SN;
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124
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125 my $histo_png = $TE_dir.'histogram.png';
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126 histogram( \%num_per_overlap_size, $histo_png, $overlap_number );
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127 print $txt "size\tnumber\tpercentage of the total overlap number\n";
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128 foreach my $k ( sort {$a <=> $b} keys %num_per_overlap_size )
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129 {
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130 my $percentage = 0;
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131 $percentage = $num_per_overlap_size{$k} * 100 / $overlap_number unless $overlap_number == 0;
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132 print $txt "$k\t$num_per_overlap_size{$k}\t"; printf $txt "%.2f\n",$percentage;
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133 }
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134 close $txt;
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135 }
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136 }
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137
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138 foreach my $tabP ( @distri_overlap )
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139 {
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140 my $sum = sum($tabP);
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141 my $ten = $tabP->[9];
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142 my $mean = mean($tabP);
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143 my $std = standard_deviation($tabP, $mean);
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144 my $zsc = z_significance($ten, $mean, $std);
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145 my $name = shift @overlaps_names;
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146 my $prob = 'NA';
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147 $prob = 1 - &Math::CDF::pnorm( $zsc ) if $zsc ne 'NA';
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148 print $ppp_f (join ("\t", $name, $sum, $ten, $mean, $std, $zsc, $prob ),"\n" );
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149 }
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150 close $ppp_f;
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151 }
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152
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153 sub count_mapped
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154 {
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155 my ( $fai, $in_file ) = @_;
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156 my ( %mapped, %dup, %has_ppp );
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157
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158 open my $f, '<', $fai || die "cannot open $fai $! \n";
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159 while(<$f>)
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160 {
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161 if ($_ =~ /(.*)\t(\d+)\n/)
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162 {
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163 $mapped{$1} = [];
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164 $mapped{$1}->[0] = []; $mapped{$1}->[1] = [];
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165 }
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166 }
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167 close $f;
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168
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169 open my $infile, "samtools view $in_file |"|| die "cannot open input file $! \n";
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170 while(<$infile>)
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171 {
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172 unless ($_ =~ /^\@[A-Za-z][A-Za-z](\t[A-Za-z][A-Za-z0-9]:[ -~]+)+$/ || $_ =~ /^\@CO\t.*/ )
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173 {
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174 my @line = split (/\t/,$_);
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175 if ($line[1] == 0)
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176 {
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177 unless ( exists ($dup{$line[3].$line[9].$line[1].$line[2]}) )
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178 {
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179 push @{$mapped{$line[2]}->[0]} , [$line[3], $line[9], $line[1], $line[2]];
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180 $has_ppp {$line[3].$line[9].$line[1].$line[2]} = 0;
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181 }
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182 $dup{$line[3].$line[9].$line[1].$line[2]}+=1;
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183 }
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184 elsif ($line[1] == 16)
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185 {
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186 unless ( exists ($dup{$line[3].$line[9].$line[1].$line[2]}) )
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187 {
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188 push @{$mapped{$line[2]}->[1]} , [$line[3], $line[9], $line[1], $line[2]];
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189 $has_ppp{$line[3].$line[9].$line[1].$line[2]} = 0;
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190 }
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191 $dup{$line[3].$line[9].$line[1].$line[2]}+=1
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192 }
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193 }
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194 }
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195 close $infile;
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196 return (\%mapped, \%dup, \%has_ppp );
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197 }
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198
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199 sub sum
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200 {
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201 my $arrayref = shift;
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202 my $result = 0;
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203 foreach (@$arrayref) {$result += $_}
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204 return $result;
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205 }
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206
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207 sub mean
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208 {
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209 my $arrayref = shift;
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210 my $result;
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211 foreach (@$arrayref) {$result += $_}
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212 return $result / scalar(@$arrayref);
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213 }
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214
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215 sub standard_deviation
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216 {
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217 my ($arrayref, $mean) = @_;
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218 return sqrt ( mean ( [map $_**2 , @$arrayref ]) - ($mean**2));
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219 }
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220
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221 sub z_significance
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222 {
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223 my ($ten, $mean, $std) = @_;
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224 my $z = 'NA';
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225 $z = (($ten - $mean) / $std) if $std != 0;
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226 return $z;
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227 }
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228
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229 1;
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230
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