package Bio::Graphics::Wiggle::Loader; =head1 SYNOPSIS my $loader = Bio::Graphics::Wiggle::Loader->new('/base/directory/for/wigfiles','wibfilename'); my $fh = IO::File->new('uploaded_file.txt'); $loader->load($fh); my $gff3_file = $loader->featurefile('gff3',$method,$source); my $featurefile = $loader->featurefile('featurefile'); my @features = $loader->features(); =head1 USAGE This module loads Bio::Graphics::Wiggle files from source files that use Jim Kent's "WIG" format: http://genome.ucsc.edu/google/goldenPath/help/wiggle.html Several data sets can be grouped together in a single WIG source file. The load() method accepts the path to a WIG source file, and will create one or more .wib ("wiggle binary") databases of quantitative data in the directory indicated when you created the loader. Call the featurefile() method to return a text file in either GFF3 or Bio::Graphics::FeatureFile format, suitable for loading into a gbrowse database. =head2 METHODS =over 4 =item $loader = Bio::Graphics::Wiggle::Loader->new('/base/directory' [,'my_data']) Create a new loader. The first argument specifies the base directory in which the loaded .wib files will be created. The second argument specifies the base name for the created .wib files, or "track" if not specified. =item $loader->load($fh) Load the data from a source WIG file opened on a filehandle. =item $data = $loader->featurefile($type [,$method,$source]) Return the data corresponding to a GFF3 or Bio::Graphics::FeatureFile. The returned file will have one feature per WIG track, and a properly formatted "wigfile" attribute that directs Bio::Graphics to the location of the quantitative data. $type is one of "gff3" or "featurefile". In the case of "gff3", you may specify an optional method and source for use in describing each feature. In the case of "featurefile", the returned file will contain GBrowse stanzas that describe a reasonable starting format to display the data. =item @features = $loader->features Returns one or more Bio::Graphics::Features objects, which can be used to create Bio::Graphics tracks with the wiggle_xyplot (and related) glyphs. =item $loader->allow_sampling(1) If allow_sampling() is passed a true value, then very large files (more than 5 MB) will undergo a sampling procedure to find their minimum and maximum values and standard deviation. Otherwise, file will be read in its entirety to generate those statistics. =back =head2 EXTENSIONS Several extensions to the WIG format "track" declaration are recognized. =over 4 =item transform= Specify a transform to be performed on all numeric data within this track prior to loading into the binary wig file. Currently, the following three declarations are recognized: transform=logtransform y' = 0 for y == 0 y' = log(y) for y > 0 y' = -log(-y) for y < 0 transform=logsquared y' = log(y**2) for y != 0 y' = 0 for y == 0 transform=none y' = y (no transform - the default) =item trim= Specify a trimming function to be performed on the data prior to scaling. Currently, the following trim functions are recognized: trim=stdev1 trim to plus/minus 1 standard deviation of the mean trim=stdev2 trim to plus/minus 2 standard deviations of the mean (default) trim=stdevN trim to plus/minus N standard deviations of the mean trim=none no trimming =back Example entended track declaration: track type=wiggle_0 name="example" description="20 degrees, 2 hr" \ trim=stdev2 transform=logsquared =cut use strict; use Carp 'croak'; use Statistics::Descriptive; use IO::Seekable; use File::Spec; use Bio::Graphics::Wiggle; use Bio::Graphics::FeatureFile; use Text::ParseWords(); use File::stat; use CGI 'escape'; use vars '%color_name'; # If a WIG file is very large (> 5 Mb) use constant BIG_FILE => 5_000_000; use constant BIG_FILE_SAMPLES => 5_000; # number of probes to make use constant DEFAULT_METHOD => 'microarray_oligo'; use constant DEFAULT_SOURCE => '.'; sub new { my $class = shift; my $base = shift or croak "Usage: Bio::Graphics::Wiggle::Loader->new('/base/path','trackname')"; my $trackname = shift || 'track'; my $wigclass = shift || 'Bio::Graphics::Wiggle'; -d $base && -w _ or croak "$base is not a writeable directory"; return bless { base => $base, tracks => {}, trackname => $trackname, tracknum => '000', track_options => {}, allow_sampling => 0, wigclass => $wigclass, },ref $class || $class; } sub allow_sampling { my $self = shift; my $d = $self->{allow_sampling}; $self->{allow_sampling} = shift if @_; $d; } sub wigclass { my $self = shift; my $d = $self->{wigclass}; $self->{wigclass} = shift if @_; return $d; } sub basedir { shift->{base} } sub wigfiles { shift->{wigfiles} } sub conf_stanzas { my $self = shift; my ($method,$source) = @_; $method ||= DEFAULT_METHOD; $source ||= DEFAULT_SOURCE; my $tracks = $self->{tracks}; my @lines = (); for my $track (sort keys %$tracks) { my $options = $tracks->{$track}{display_options}; my $name = $options->{name} ||= $track; $options->{visibility} ||= 'dense'; $options->{color} ||= $options->{visibility} =~ /pack/i ? '255,0,0' : '0,0,0'; $options->{altColor} ||= $options->{visibility} =~ /pack/i ? '0,0,255' : '0,0,0'; # stanza push @lines,"[$track]"; if (my $graph_type = $options->{glyph}) { if ($graph_type =~ /box/) { push @lines, "glyph = wiggle_box"; } else { push @lines,"glyph = ". ($graph_type =~/density/ ? 'wiggle_density' : 'wiggle_xyplot'); } } else { push @lines,"glyph = ". ($options->{visibility}=~/pack/ ? 'wiggle_density' : 'wiggle_xyplot'); } push @lines,"key = $options->{name}" if $options->{name}; push @lines,"description = $options->{description}" if $options->{description}; if (my $color = $options->{color}) { push @lines,"bgcolor=".format_color($color); } if (my $color = $options->{altColor}) { push @lines,"fgcolor=" . format_color($color); } if (exists $options->{viewLimits} and my ($low,$hi) = split ':',$options->{viewLimits}) { push @lines,"min_score = $low"; push @lines,"max_score = $hi"; } if (exists $options->{maxHeightPixels} and my ($max,$default,$min) = split ':',$options->{maxHeightPixels}) { push @lines,"height = $default"; } push @lines,"smoothing = $options->{windowingFunction}" if $options->{windowingFunction}; my $smoothing_window = $options->{smoothingWindow} || 0; push @lines,"smoothing window = $options->{smoothingWindow}" if $options->{smoothingWindow}; push @lines,''; } return join "\n",@lines; } sub featurefile { my $self = shift; my $type = shift; my ($method,$source) = @_; $method ||= DEFAULT_METHOD; $source ||= DEFAULT_SOURCE; $type ||= 'featurefile'; $type =~ /^(gff3|featurefile)$/i or croak "featurefile type must be one of 'gff3' or 'featurefile'"; my @lines; my $tracks = $self->{tracks}; if ($type eq 'gff3') { push @lines,"##gff-version 3",""; } else { push @lines,$self->conf_stanzas($method,$source),""; } for my $track (sort keys %$tracks) { my $options = $tracks->{$track}{display_options}; my $name = $options->{name} ||= $track; my $seqids = $tracks->{$track}{seqids}; my $note = escape($options->{description}); my @attributes; push @attributes,qq(Name=$name) if defined $name; push @attributes,qq(Note=$note) if defined $note; # data, sorted by chromosome my @seqid = sort keys %$seqids; for my $seqid (@seqid) { $seqid or next; $tracks->{$track}{seqids}{$seqid}{wig}->write(); my $attributes = join ';',(@attributes,"wigfile=$seqids->{$seqid}{wigpath}"); if ($type eq 'gff3') { push @lines,join "\t",($seqid,$source,$method, $seqids->{$seqid}{start}, $seqids->{$seqid}{end}, '.','.','.', $attributes ); } else { push @lines,''; push @lines,"reference=$seqid"; push @lines,"$track $seqid.data $seqids->{$seqid}{start}..$seqids->{$seqid}{end} $attributes"; } } } return join("\n",@lines)."\n"; } sub features { my $self = shift; my $text = $self->featurefile('featurefile'); my $file = Bio::Graphics::FeatureFile->new(-text=>$text); return $file->features; } sub load { my $self = shift; my $infh = shift; my $format = 'none'; local $_; LINE: while (<$infh>) { chomp; next if /^#/; next unless /\S/; if (/^track/) { $self->process_track_line($_); next; } if (/^fixedStep/) { $self->process_fixed_step_declaration($_); $format = 'fixed'; } if (/^variableStep/) { $self->process_variable_step_declaration($_); $format = 'variable'; } if (/^\S+\s+\d+\s+\d+\s+-?[\dEe.]+/) { $self->process_first_bedline($_); $format = 'bed'; } if ($format ne 'none') { # remember where we are, find min and max values, return my $pos = tell($infh); $self->minmax($infh,$format eq 'bed' ? $_ : '') unless $self->{track_options}{chrom} && exists $self->current_track->{seqids}{$self->{track_options}{chrom}}{min}; seek($infh,$pos,0); $self->process_bed($infh,$_) if $format eq 'bed'; $self->process_fixedline($infh) if $format eq 'fixed'; $self->process_variableline($infh) if $format eq 'variable'; $format = 'none'; } redo LINE if defined $_ && /^(track|variableStep|fixedStep)/; } return 1; } sub process_track_line { my $self = shift; my $line = shift; my @tokens = shellwords($line); shift @tokens; my %options = map {split '='} @tokens; $options{type} eq 'wiggle_0' or croak "invalid/unknown wiggle track type $options{type}"; delete $options{type}; $self->{tracknum}++; $self->current_track->{display_options} = \%options; } sub process_fixed_step_declaration { my $self = shift; my $line = shift; my @tokens = shellwords($line); shift @tokens; my %options = map {split '='} @tokens; exists $options{chrom} or croak "invalid fixedStep line: need a chrom option"; exists $options{start} or croak "invalid fixedStep line: need a start option"; exists $options{step} or croak "invalid fixedStep line: need a step option"; $self->{track_options} = \%options; } sub process_variable_step_declaration { my $self = shift; my $line = shift; my @tokens = shellwords($line); shift @tokens; my %options = map {split '='} @tokens; exists $options{chrom} or croak "invalid variableStep line: need a chrom option"; $self->{track_options} = \%options; } sub process_first_bedline { my $self = shift; my $line = shift; my @tokens = shellwords($line); $self->{track_options} = {chrom => $tokens[0]}; } sub current_track { my $self = shift; return $self->{tracks}{$self->{tracknum}} ||= {}; } sub minmax { my $self = shift; my ($infh,$bedline) = @_; local $_; my $transform = $self->get_transform; my $seqids = ($self->current_track->{seqids} ||= {}); my $chrom = $self->{track_options}{chrom}; if ($self->allow_sampling && (my $size = stat($infh)->size) > BIG_FILE) { warn "Wiggle file is very large; resorting to genome-wide sample statistics for $chrom.\n"; $self->{FILEWIDE_STATS} ||= $self->sample_file($infh,BIG_FILE_SAMPLES); for (keys %{$self->{FILEWIDE_STATS}}) { $seqids->{$chrom}{$_} = $self->{FILEWIDE_STATS}{$_}; } return; } my %stats; if ($bedline) { # left-over BED line my @tokens = split /\s+/,$bedline; my $seqid = $tokens[0]; my $value = $tokens[-1]; $value = $transform->($self,$value) if $transform; $stats{$seqid} ||= Statistics::Descriptive::Sparse->new(); $stats{$seqid}->add_data($value); } while (<$infh>) { last if /^track/; last if /chrom=(\S+)/ && $1 ne $chrom; next if /^\#|fixedStep|variableStep/; my @tokens = split(/\s+/,$_) or next; my $seqid = @tokens > 3 ? $tokens[0] : $chrom; my $value = $tokens[-1]; $value = $transform->($self,$value) if $transform; $stats{$seqid} ||= Statistics::Descriptive::Sparse->new(); $stats{$seqid}->add_data($value); } for my $seqid (keys %stats) { $seqids->{$seqid}{min} = $stats{$seqid}->min(); $seqids->{$seqid}{max} = $stats{$seqid}->max(); $seqids->{$seqid}{mean} = $stats{$seqid}->mean(); $seqids->{$seqid}{stdev} = $stats{$seqid}->standard_deviation(); } } sub sample_file { my $self = shift; my ($fh,$samples) = @_; my $transform = $self->get_transform; my $stats = Statistics::Descriptive::Sparse->new(); my $size = stat($fh)->size; my $count=0; while ($count < $samples) { seek($fh,int(rand $size),0) or die; scalar <$fh>; # toss first line my $line = <$fh>; # next full line $line or next; my @tokens = split /\s+/,$line; my $value = $tokens[-1]; next unless $value =~ /^[\d\seE.+-]+$/; # non-numeric $value = $transform->($self,$value) if $transform; $stats->add_data($value); $count++; } return { min => $stats->min, max => $stats->max, mean => $stats->mean, stdev => $stats->standard_deviation, }; } sub get_transform { my $self = shift; my $transform = $self->current_track->{display_options}{transform}; return $self->can($transform) if $transform; } # one and only transform currently defined # Natural log of the square of the value. # Return 0 if the value is 0 sub logsquared { my $self = shift; my $value = shift; return 0 if $value == 0; return log($value**2); } sub logtransform { my $self = shift; my $value = shift; return 0 if $value == 0; if ($value < 0) { return -log(-$value); } else { return log($value); } } sub process_bed { my $self = shift; my $infh = shift; my $oops = shift; my $transform = $self->get_transform; $self->process_bedline($oops) if $oops; while (<$infh>) { last if /^track/; next if /^#/; chomp; $self->process_bedline($_); } } sub process_bedline { my $self = shift; my ($line,$transform) = @_; my ($seqid,$start,$end,$value) = split /\s+/,$line; $value = $transform->($self,$value) if $transform; $start++; # to 1-based coordinates my $wigfile = $self->wigfile($seqid); $wigfile->set_range($start=>$end, $value); # update span $self->current_track->{seqids}{$seqid}{start} = $start unless exists $self->current_track->{seqids}{$seqid}{start} and $self->current_track->{seqids}{$seqid}{start} < $start; $self->current_track->{seqids}{$seqid}{end} = $end unless exists $self->current_track->{seqids}{$seqid}{end} and $self->current_track->{seqids}{$seqid}{end} > $end; } sub process_fixedline { my $self = shift; my $infh = shift; my $seqid = $self->{track_options}{chrom}; my $wigfile = $self->wigfile($seqid); my $start = $self->{track_options}{start}; my $step = $self->{track_options}{step}; my $span = $wigfile->span; # update start and end positions $self->{track_options}{span} ||= $wigfile->span || 1; my $chrom = $self->current_track->{seqids}{$seqid}; $chrom->{start} = $start if !defined $chrom->{start} || $chrom->{start} > $start; my $end = $chrom->{start} + $span - 1; $chrom->{end} = $end if !defined $chrom->{end} || $chrom->{end} < $end; my $transform = $self->get_transform; # write out data in 500K chunks for efficiency my @buffer; while (<$infh>) { last if /^(track|variableStep|fixedStep)/; next if /^#/; chomp; push @buffer,$_; if (@buffer >= 500_000) { @buffer = map {$transform->($self,$_)} @buffer if $transform; $wigfile->set_values($start=>\@buffer); my $big_step = $step * @buffer; $start += $big_step; $self->current_track->{seqids}{$seqid}{end} = $start + $big_step - 1 + $span; @buffer = (); # reset at the end } } @buffer = map {$transform->($self,$_)} @buffer if $transform; $wigfile->set_values($start=>\@buffer) if @buffer; $self->current_track->{seqids}{$seqid}{end} = $start + @buffer*$step - 1 + $span; } sub process_variableline { my $self = shift; my $infh = shift; my $seqid = $self->{track_options}{chrom}; my $chrom = $self->current_track->{seqids}{$seqid}; my $wigfile = $self->wigfile($seqid); my $span = $wigfile->span; my $transform = $self->get_transform; while (<$infh>) { last if /^(track|variableStep|fixedStep)/; next if /^#/; chomp; my ($start,$value) = split /\s+/ or next; $value = $transform->($self,$value) if $transform; eval { $wigfile->set_value($start=>$value); 1; } or croak "Data error on line $.: $_\nDetails: $@"; # update span $chrom->{start} = $start if !defined $chrom->{start} || $chrom->{start} > $start; my $end = $start + $span - 1; $chrom->{end} = $end if !defined $chrom->{end} || $chrom->{end} < $end; } $self->current_track->{seqids}{$seqid}{end} ||= $self->current_track->{seqids}{$seqid}{start}; } sub wigfile { my $self = shift; my $seqid = shift; my $ts = time(); my $current_track = $self->{tracknum}; my $tname = $self->{trackname} || 'track'; unless (exists $self->current_track->{seqids}{$seqid}{wig}) { my $path = File::Spec->catfile($self->{base},"$tname\_$current_track.$seqid.$ts.wib"); my @stats; foreach (qw(min max mean stdev)) { my $value = $self->current_track->{seqids}{$seqid}{$_} || $self->{FILEWIDE_STATS}{$_} || next; push @stats,($_=>$value); } my $step = $self->{track_options}{step} || 1; my $span = $self->{track_options}{span} || $self->{track_options}{step} || 1; my $trim = $self->current_track->{display_options}{trim} || 'stdev10'; my $transform = $self->current_track->{display_options}{transform}; my $class = $self->wigclass; unless ($class->can('new')) { warn "loading $class"; eval "require $class"; die $@ if $@; } my $wigfile = $class->new( $path, 1, { seqid => $seqid, step => $step, span => $span, trim => $trim, @stats, }, ); $wigfile or croak "Couldn't create wigfile $wigfile: $!"; $self->current_track->{seqids}{$seqid}{wig} = $wigfile; $self->current_track->{seqids}{$seqid}{wigpath} = $path; } return $self->current_track->{seqids}{$seqid}{wig}; } sub format_color { my $rgb = shift; return $rgb unless $rgb =~ /\d+,\d+,\d+/; my ($r,$g,$b) = split ',',$rgb; my $hex = '#'.join '',map {sprintf("%02X",$_)}($r,$g,$b); return translate_color($hex); } # use English names for the most common colors sub translate_color { my $clr = shift; unless (%color_name) { while () { chomp; my ($hex,$name) = split or next; $color_name{$hex} = $name; } } return $color_name{$clr} || $clr; } # work around an annoying uninit variable warning from Text::Parsewords sub shellwords { my @args = @_; return unless @args; foreach(@args) { s/^\s+//; s/\s+$//; $_ = '' unless defined $_; } my @result = Text::ParseWords::shellwords(@args); return @result; } 1; __DATA__ #000000 black #FFFFFF white #0000FF blue #00FF00 green #FF0000 red #FFFF00 yellow #00FFFF cyan #FF00FF magenta #C0C0C0 gray __END__ =head1 SEE ALSO L, L, L, L, L, L, L =head1 AUTHOR Lincoln Stein Elstein@cshl.orgE. Copyright (c) 2007 Cold Spring Harbor Laboratory This package and its accompanying libraries is free software; you can redistribute it and/or modify it under the terms of the GPL (either version 1, or at your option, any later version) or the Artistic License 2.0. Refer to LICENSE for the full license text. In addition, please see DISCLAIMER.txt for disclaimers of warranty. =cut