# $Id: ExtendedSignalp.pm 16123 2009-09-17 12:57:27Z cjfields $ # # BioPerl module for Bio::Tools::Signalp::ExtendedSignalp # # Please direct questions and support issues to # # Cared for by Emmanuel Quevillon # # Copyright Emmanuel Quevillon # # You may distribute this module under the same terms as perl itself # # POD documentation - main docs before the code =head1 NAME Bio::Tools::Signalp::ExtendedSignalp - enhanced parser for Signalp output =head1 SYNOPSIS use Bio::Tools::Signalp::ExtendedSignalp; my $params = [qw(maxC maxY maxS meanS D)]; my $parser = new Bio::Tools::Signalp::ExtendedSignalp( -fh => $filehandle -factors => $params ); $parser->factors($params); while( my $sp_feat = $parser->next_feature ) { #do something #eg push @sp_feat, $sp_feat; } =head1 DESCRIPTION # Please direct questions and support issues to I Parser module for Signalp. Based on the EnsEMBL module Bio::EnsEMBL::Pipeline::Runnable::Protein::Signalp originally written by Marc Sohrmann (ms2 a sanger.ac.uk) Written in BioPipe by Balamurugan Kumarasamy (savikalpa a fugu-sg.org) Cared for by the Fugu Informatics team (fuguteam@fugu-sg.org) You may distribute this module under the same terms as perl itself Compared to the original SignalP, this method allow the user to filter results out based on maxC maxY maxS meanS and D factor cutoff for the Neural Network (NN) method only. The HMM method does not give any filters with 'YES' or 'NO' as result. The user must be aware that the filters can only by applied on NN method. Also, to ensure the compability with original Signalp parsing module, the user must know that by default, if filters are empty, max Y and mean S filters are automatically used to filter results. If the used gives a list, then the parser will only report protein having 'YES' for each factor. This module supports parsing for full, summary and short output form signalp. Actually, full and summary are equivalent in terms of filtering results. =head1 FEEDBACK =head2 Mailing Lists User feedback is an integral part of the evolution of this and other Bioperl modules. Send your comments and suggestions preferably to the Bioperl mailing list. Your participation is much appreciated. bioperl-l@bioperl.org - General discussion http://bioperl.org/wiki/Mailing_lists - About the mailing lists =head2 Support Please direct usage questions or support issues to the mailing list: I rather than to the module maintainer directly. Many experienced and reponsive experts will be able look at the problem and quickly address it. Please include a thorough description of the problem with code and data examples if at all possible. =head2 Reporting Bugs Report bugs to the Bioperl bug tracking system to help us keep track of the bugs and their resolution. Bug reports can be submitted via the web: http://bugzilla.open-bio.org/ =head1 AUTHOR Based on the Bio::Tools::Signalp module Emmanuel Quevillon =head1 APPENDIX The rest of the documentation details each of the object methods. Internal methods are usually preceded with a _ =cut package Bio::Tools::Signalp::ExtendedSignalp; use strict; use Data::Dumper; use Bio::SeqFeature::Generic; # don't need Bio::Root::Root/IO (already in inheritance tree) use base qw(Bio::Tools::Signalp Bio::Tools::AnalysisResult); #Supported arguments my $FACTS = { 'maxC' => 1, 'maxS' => 1, 'maxY' => 1, 'meanS' => 1, 'D' => 1, }; =head2 new Title : new Usage : my $obj = new Bio::Tools::Signalp::ExtendedSignalp(); Function: Builds a new Bio::Tools::Signalp::ExtendedSignalp object Returns : Bio::Tools::Signalp::ExtendedSignalp Args : -fh/-file => $val, # for initing input, see Bio::Root::IO =cut sub new { my($class,@args) = @_; my $self = $class->SUPER::new(@args); $self->_initialize_io(@args); my $factors = $self->_rearrange([qw(FACTORS)], @args); #To behave like the parent module (Bio::Tools::Signalp) we default factors to these two factors if($factors && scalar(@$factors)){ $factors = $factors; } else{ $factors = [qw(maxY meanS)]; } $factors && $self->factors($factors); return $self; } =head2 next_feature Title : next_feature Usage : my $feat = $signalp->next_feature Function: Get the next result feature from parser data Returns : Bio::SeqFeature::Generic Args : none =cut sub next_feature { my ($self) = @_; if(!$self->_parsed()){ $self->_parse(); } return shift @{$self->{_features}} || undef; } =head2 _filterok Title : _filterok Usage : my $feat = $signalp->_filterok Function: Check if the factors required by the user are all ok. Returns : 1/0 Args : hash reference =cut sub _filterok { my($self, $hash) = @_; #We hope everything will be fine ;) my $bool = 1; #If the user did not give any filter, we keep eveything return $bool unless keys %{$self->{_factors}}; #If only one of the factors parsed is equal to NO based on the user factors cutoff #Then the filter is not ok. foreach my $fact (keys %{$self->factors()}){ if(exists($hash->{$fact}) && $hash->{$fact} =~ /^N/){ $bool = 0; } } return $bool; } =head2 factors Title : factors Usage : my $feat = $signalp->factors Function: Get/Set the filters required from the user Returns : hash Args : array reference =cut sub factors { my($self, $array) = @_; if($array){ $self->{_factors} = { }; foreach my $f (@$array){ if(exists($FACTS->{$f})){ $self->{_factors}->{$f} = 1; } else{ $self->throw("[$f] incorrect factor. Supported:\n- ".join("\n- ", keys %$FACTS)."\n"); } } } return $self->{_factors}; } =head2 _parsed Title : _parsed Usage : obj->_parsed() Function: Get/Set if the result is parsed or not Returns : 1/0 scalar Args : On set 1 =cut sub _parsed { my($self, $parsed) = @_; if(defined($parsed)){ $self->{_parsed} = $parsed; } return $self->{_parsed}; } =head2 _parse Title : _parse Usage : obj->_parse Function: Parse the SignalP result Returns : Args : =cut sub _parse { my($self) = @_; #Let's read the file... while (my $line = $self->_readline()) { chomp $line; #We want to be sure to catch the first non empty line to be ablte to determine #which format we are working with... next unless ($line =~ /^>(\S+)|^# SignalP-[NHM]+ \S+ predictions/); if($line =~ /^>(\S+)/){ $self->_pushback($line); $self->_parse_summary_format(); last; } elsif($line =~ /^# SignalP-[NHM]+ \S+ predictions/){ $self->_pushback($line); $self->_parse_short_format(); last; } else{ $self->throw("Unable to determine the format type."); } } return; } =head2 _parse_summary_format Title : _parse_summary_format Usage : $self->_parse_summary_format Function: Method to parse summary/full format from signalp output It automatically fills filtered features. Returns : Args : =cut sub _parse_summary_format { my($self) = @_; my $feature = undef; my $ok = 0; while(my $line = $self->_readline()){ if($line =~ /^SignalP-NN result:/){ $self->_pushback($line); $feature = $self->_parse_nn_result($feature); } if($line =~ /^SignalP-HMM result:/){ $self->_pushback($line); $feature = $self->_parse_hmm_result($feature); } if($line =~ /^---------/ && $feature){ my $new_feature = $self->create_feature($feature); push @{$self->{_features}}, $new_feature if $new_feature; $feature = undef; } } return; } =head2 _parse_nn_result Title : _parse_nn_result Usage : obj->_parse_nn_result Function: Parses the Neuronal Network (NN) part of the result Returns : Hash reference Args : =cut sub _parse_nn_result { my($self, $feature) = @_; my $ok = 0; my %facts; #SignalP-NN result: #>MGG_11635.5 length = 100 ## Measure Position Value Cutoff signal peptide? # max. C 37 0.087 0.32 NO # max. Y 37 0.042 0.33 NO # max. S 3 0.062 0.87 NO # mean S 1-36 0.024 0.48 NO # D 1-36 0.033 0.43 NO while(my $line = $self->_readline()){ chomp $line; if($line =~ /^SignalP-NN result:/){ $ok = 1; next; } $self->throw("Wrong line for parsing NN results.") unless $ok; if ($line=~/^\>(\S+)\s+length/) { $self->seqname($1); %facts = (); next; } elsif($line =~ /max\.\s+C\s+(\S+)\s+\S+\s+\S+\s+(\S+)/) { $feature->{maxCprob} = $1; $facts{maxC} = $2; next; } elsif ($line =~ /max\.\s+Y\s+(\S+)\s+\S+\s+\S+\s+(\S+)/) { $feature->{maxYprob} = $1; $facts{maxY} = $2; next; } elsif($line =~ /max\.\s+S\s+(\S+)\s+\S+\s+\S+\s+(\S+)/) { $feature->{maxSprob} = $1; $facts{maxS} = $2; next; } elsif ($line=~/mean\s+S\s+(\S+)\s+\S+\s+\S+\s+(\S+)/) { $feature->{meanSprob} = $1; $facts{meanS} = $2; next; } elsif ($line=~/\s+D\s+(\S+)\s+\S+\s+\S+\s+(\S+)/) { $feature->{Dprob} = $1; $facts{D} = $2; next; } #If we don't have this line it means that all the factors cutoff are equal to 'NO' elsif ($line =~ /Most likely cleavage site between pos\.\s+(\d+)/) { #if($self->_filterok(\%facts)){ #$feature->{name} = $self->seqname(); #$feature->{start} = 1; $feature->{end} = $1 + 1; #To be consistent with end given in short format #} #return $feature; } elsif($line =~ /^\s*$/){ last; } } if($self->_filterok(\%facts)){ $feature->{name} = $self->seqname(); $feature->{start} = 1; $feature->{nnPrediction} = 'signal-peptide'; } return $feature; } =head2 _parse_hmm_result Title : _parse_hmm_result Usage : obj->_parse_hmm_result Function: Parses the Hiden Markov Model (HMM) part of the result Returns : Hash reference Args : =cut sub _parse_hmm_result { my ($self, $feature_hash) = @_; my $ok = 0; #SignalP-HMM result: #>MGG_11635.5 #Prediction: Non-secretory protein #Signal peptide probability: 0.000 #Signal anchor probability: 0.000 #Max cleavage site probability: 0.000 between pos. -1 and 0 while(my $line = $self->_readline()){ chomp $line; next if $line =~ /^\s*$/o; if($line =~ /^SignalP-HMM result:/){ $ok = 1; next; } $self->throw("Wrong line for parsing HMM result.") unless $ok; if($line =~ /^>(\S+)/){ #In case we already seen a name with NN results $feature_hash->{name} = $1 unless $self->seqname(); } elsif($line =~ /Prediction: (.+)$/){ $feature_hash->{hmmPrediction} = $1; } elsif($line =~ /Signal peptide probability: ([0-9\.]+)/){ $feature_hash->{peptideProb} = $1; } elsif($line =~ /Signal anchor probability: ([0-9\.]+)/){ $feature_hash->{anchorProb} = $1; } elsif($line =~ /Max cleavage site probability: (\S+) between pos. \S+ and (\S+)/){ $feature_hash->{cleavageSiteProb} = $1; #Strange case, if we don't have an end value in NN result (no nn method launched) #We try anyway to get an end value, unless this value is lower than 1 which is #the start $feature_hash->{end} = $2 if($2 > 1 && !$feature_hash->{end}); $feature_hash->{start} = 1 unless $feature_hash->{start}; last; } } return $feature_hash; } =head2 _parse_short_format Title : _parse_short_format Usage : $self->_parse_short_format Function: Method to parse short format from signalp output It automatically fills filtered features. Returns : Args : =cut sub _parse_short_format { my($self) = @_; my $ok = 0; my $method = undef; $self->{_oformat} = 'short'; #Output example # SignalP-NN euk predictions # SignalP-HMM euk predictions # name Cmax pos ? Ymax pos ? Smax pos ? Smean ? D ? # name ! Cmax pos ? Sprob ? #Q5A8M1_CANAL 0.085 27 N 0.190 35 N 0.936 27 Y 0.418 N 0.304 N Q5A8M1_CANAL Q 0.001 35 N 0.002 N #O74127_YARLI 0.121 21 N 0.284 21 N 0.953 11 Y 0.826 Y 0.555 Y O74127_YARLI S 0.485 23 N 0.668 Y #Q5VJ86_9PEZI 0.355 24 Y 0.375 24 Y 0.798 12 N 0.447 N 0.411 N Q5VJ86_9PEZI Q 0.180 23 N 0.339 N #Q5A8U5_CANAL 0.085 27 N 0.190 35 N 0.936 27 Y 0.418 N 0.304 N Q5A8U5_CANAL Q 0.001 35 N 0.002 N while(my $line = $self->_readline()){ chomp $line; next if $line =~ /^\s*$|^# name/; if($line =~ /^#/){ $method = $line =~ /SignalP-NN .+ SignalP-HMM/ ? 'both' : $line =~ /SignalP-NN/ ? 'nn' : 'hmm'; next; } #$self->throw("It looks like the format is not 'short' format.") unless($ok); my @data = split(/\s+/, $line); $self->seqname($data[0]); my $factors = { }; my $feature = { }; #NN results gives more fields than HMM if($method eq 'both' || $method eq 'nn'){ $feature->{maxCprob} = $data[1]; $factors->{maxC} = $data[3]; $feature->{maxYprob} = $data[4]; $factors->{maxY} = $data[6]; $feature->{maxSprob} = $data[7]; $factors->{maxS} = $data[9]; $feature->{meanSprob}= $data[10]; $factors->{meanS} = $data[11]; $feature->{Dprob} = $data[12]; $factors->{D} = $data[13]; #It looks like the max Y position is reported as the most likely cleavage position $feature->{end} = $data[5]; $feature->{nnPrediction} = 'signal-peptide'; if($method eq 'both'){ $feature->{hmmPrediction} = $data[15] eq 'Q' ? 'Non-secretory protein' : 'Signal peptide'; $feature->{cleavageSiteProb} = $data[16]; $feature->{peptideProb} = $data[19]; } } elsif($method eq 'hmm'){ #In short output anchor probability is not given $feature->{hmmPrediction} = $data[1] eq 'Q' ? 'Non-secretory protein' : 'Signal peptide'; $feature->{cleavageSiteProb} = $data[2]; $feature->{peptideProb} = $data[5]; #It looks like the max cleavage probability position is given by the Cmax proability $feature->{end} = $data[3]; } #Unfortunately, we cannot parse the filters for hmm method. if($self->_filterok($factors)){ $feature->{name} = $self->seqname(); $feature->{start} = 1; $feature->{source} = 'Signalp'; $feature->{primary} = 'signal_peptide'; $feature->{program} = 'Signalp'; $feature->{logic_name} = 'signal_peptide'; my $new_feat = $self->create_feature($feature); push @{$self->{_features}}, $new_feat if $new_feat; } } return; } =head2 create_feature Title : create_feature Usage : obj->create_feature(\%feature) Function: Internal(not to be used directly) Returns : Args : =cut sub create_feature { my ($self, $feat) = @_; #If we don't have neither start nor end, we return. unless($feat->{name} && $feat->{start} && $feat->{end}){ return; } # create feature object my $feature = Bio::SeqFeature::Generic->new( -seq_id => $feat->{name}, -start => $feat->{start}, -end => $feat->{end}, -score => defined($feat->{peptideProb}) ? $feat->{peptideProb} : '', -source => 'Signalp', -primary => 'signal_peptide', -logic_name => 'signal_peptide', ); $feature->add_tag_value('peptideProb', $feat->{peptideProb}); $feature->add_tag_value('anchorProb', $feat->{anchorProb}); $feature->add_tag_value('evalue',$feat->{anchorProb}); $feature->add_tag_value('percent_id','NULL'); $feature->add_tag_value("hid",$feat->{primary}); $feature->add_tag_value('signalpPrediction', $feat->{hmmPrediction}); $feature->add_tag_value('cleavageSiteProb', $feat->{cleavageSiteProb}) if($feat->{cleavageSiteProb}); $feature->add_tag_value('nnPrediction', $feat->{nnPrediction}) if($feat->{nnPrediction}); $feature->add_tag_value('maxCprob', $feat->{maxCprob}) if(defined($feat->{maxCprob})); $feature->add_tag_value('maxSprob', $feat->{maxSprob}) if(defined($feat->{maxSprob})); $feature->add_tag_value('maxYprob', $feat->{maxYprob}) if(defined($feat->{maxYprob})); $feature->add_tag_value('meanSprob', $feat->{meanSprob}) if(defined($feat->{meanSprob})); $feature->add_tag_value('Dprob', $feat->{Dprob}) if(defined($feat->{Dprob})); return $feature; } =head2 seqname Title : seqname Usage : obj->seqname($name) Function: Internal(not to be used directly) Returns : Args : =cut sub seqname{ my ($self,$seqname)=@_; if (defined($seqname)){ $self->{'seqname'} = $seqname; } return $self->{'seqname'}; } 1;