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#
# BioPerl module for Bio::Tools::Run::Phylo::Phast::PhastCons
#
# Please direct questions and support issues to <bioperl-l@bioperl.org>
#
# Cared for by Sendu Bala <bix@sendu.me.uk>
#
# Copyright Sendu Bala
#
# You may distribute this module under the same terms as perl itself
# POD documentation - main docs before the code
=head1 NAME
Bio::Tools::Run::Phylo::Phast::PhastCons - Wrapper for footprinting using
phastCons
=head1 SYNOPSIS
use Bio::Tools::Run::Phylo::Phast::PhastCons;
# Make a PhastCons factory
$factory = Bio::Tools::Run::Phylo::Phast::PhastCons->new();
# Pass the factory an alignment and the corresponding species tree
$align_filename = 't/data/apes.multi_fasta';
$species_tree_filename = 't/data/apes.newick';
@features = $factory->run($align_filename, $species_tree_filename);
# or get a Bio::Align::AlignI (SimpleAlign) object from somewhere, and
# generate the species tree automatically using a Bio::DB::Taxonomy database
$tdb = Bio::DB::Taxonomy->new(-source => 'entrez');
@features = $factory->run($aln_obj, $tdb);
# @features is an array of Bio::SeqFeature::Annotated, one feature per
# alignment sequence and prediction
=head1 DESCRIPTION
This is a wrapper for running the phastCons application by Adam Siepel. You
can get details here: http://compgen.bscb.cornell.edu/~acs/software.html
phastCons is used for phylogenetic footprinting/ shadowing.
Currently the interface is extremely simplified, allowing only one
analysis method. The focus here is on ease of use, allowing phastCons
to estimate as many parameters as possible and having it output just
the 'most conserved' blocks it detects. You can, however, try
supplying normal phastCons arguments to new(), or calling arg-named
methods (excluding initial hyphens and converting others to
underscores, eg. $factory-E<gt>indels_only(1) to set the --indels-only
arg).
The particular analysis carried out here is to:
1. Use phyloFit to generate a tree model for initialization of the nonconserved
model from the supplied alignment (all data) and species tree
2. Run phastCons in 'training' mode for parameter estimation using all the
alignment data and the model from step 1
3. Run phastCons with the trees from step 2 to discover the most conserved
regions
See the 'HowTo' at http://compgen.bscb.cornell.edu/~acs/phastCons-HOWTO.html
for details on how to improve results.
WARNING: the API is likely to change in the future to allow for alternative
analysis types.
You will need to enable this phastCons wrapper to find the phast programs (at
least phastCons and phyloFit).
This can be done in (at least) three ways:
1. Make sure the phastCons and phyloFit executables are in your path.
2. Define an environmental variable PHASTDIR which is a
directory which contains the phastCons and phyloFit applications:
In bash:
export PHASTDIR=/home/username/phast/bin
In csh/tcsh:
setenv PHASTDIR /home/username/phast/bin
3. Include a definition of an environmental variable PHASTDIR in
every script that will use this PhastCons wrapper module, e.g.:
BEGIN { $ENV{PHASTDIR} = '/home/username/phast/bin' }
use Bio::Tools::Run::Phylo::Phast::PhastCons;
=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<bioperl-l@bioperl.org>
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://redmine.open-bio.org/projects/bioperl/
=head1 AUTHOR - Sendu Bala
Email bix@sendu.me.uk
=head1 APPENDIX
The rest of the documentation details each of the object methods.
Internal methods are usually preceded with a _
=cut
package Bio::Tools::Run::Phylo::Phast::PhastCons;
use strict;
use Cwd;
use File::Basename;
use Clone qw(clone);
use Bio::AlignIO;
use Bio::Tools::Run::Phylo::Phast::PhyloFit;
use Bio::FeatureIO;
use Bio::Annotation::SimpleValue;
use base qw(Bio::Tools::Run::Phylo::PhyloBase);
our $PROGRAM_NAME = 'phastCons';
our $PROGRAM_DIR = $ENV{'PHASTDIR'};
# methods and their synonyms from the phastCons args we support
our %PARAMS = (rho => 'R',
nrates => 'k',
transitions => 't',
target_coverage => 'C',
expected_length => ['E', 'expected_lengths'],
lnl => 'L',
log => 'g',
max_micro_indel => 'Y',
indel_params => 'D',
lambda => 'l',
extrapolate => 'e',
hmm => 'H',
catmap => 'c',
states => 'S',
reflect_strand => 'U',
require_informative => 'M',
not_informative => 'F');
our %SWITCHES = (quiet => 'q',
indels => 'I',
indels_only => 'J',
FC => 'X',
coding_potential => 'p',
ignore_missing => 'z');
# just to be explicit, args we don't support (yet) or we handle ourselves
our %UNSUPPORTED = (estimate_trees => 'T',
estimate_rho => 'O',
gc => 'G',
msa_format => 'i',
score => 's',
no_post_probs => 'n',
seqname => 'N',
refidx => 'r',
idpref => 'P',
help => 'h',
alias => 'A',
most_conserved => ['V', 'viterbi']);
=head2 program_name
Title : program_name
Usage : $factory>program_name()
Function: holds the program name
Returns : string
Args : None
=cut
sub program_name {
return $PROGRAM_NAME;
}
=head2 program_dir
Title : program_dir
Usage : $factory->program_dir(@params)
Function: returns the program directory, obtained from ENV variable.
Returns : string
Args : None
=cut
sub program_dir {
return $PROGRAM_DIR;
}
=head2 new
Title : new
Usage : $factory = Bio::Tools::Run::Phylo::Phast::PhastCons->new(@params)
Function: Creates a new PhastCons factory
Returns : Bio::Tools::Run::Phylo::Phast::PhastCons
Args : Optionally, provide any of the following (defaults are not to use,
see the same-named methods for information on what each option does):
{
-target_coverage => number between 0 and 1
AND
-expected_length => int
}
-rho => number between 0 and 1
-quiet => boolean (turn on or off program output to console)
Most other options understood by phastCons can be supplied as key =>
value pairs in this way. Options that don't normally take a value
should be given a value of 1. You can type the keys as you would on
the command line (eg. '--indels-only' => 1) or with only a single
hyphen to start and internal hyphens converted to underscores (eg.
-indels_only => 1) to avoid having to quote the key.
These options can NOT be used with this wrapper currently:
estimate_trees / T
estimate_rho / O
gc / G
msa_format / i
score / s
no_post_probs / n
seqname / N
idpref / P
help / h
alias / A
most_conserved / V / viterbi
refidx / r
=cut
sub new {
my ($class, @args) = @_;
my $self = $class->SUPER::new(@args);
$self->_set_from_args(\@args, -methods => {(map { $_ => $PARAMS{$_} } keys %PARAMS),
(map { $_ => $SWITCHES{$_} } keys %SWITCHES)},
-create => 1);
return $self;
}
=head2 target_coverage
Title : target_coverage
Usage : $factory->target_coverage(0.25);
Function: Constrain transition parameters such that the expected fraction of
sites in conserved elements is the supplied value.
Returns : number (default undef)
Args : None to get, number (between 0 and 1) to set
=cut
sub target_coverage {
my ($self, $num) = @_;
if (defined ($num)) {
($num > 0 && $num < 1) || $self->throw("target_coverage value must be between 0 and 1, exclusive");
$self->{coverage} = $num;
}
return $self->{coverage} || return;
}
=head2 expected_length
Title : expected_length
Usage : $factory->expected_length(5);
Function: Set transition probabilities such that the expected length of a
conserved element is the supplied value. target_coverage() must also
be set.
Returns : int (default undef)
Args : None to get, int to set
=cut
# created automatically
=head2 rho
Title : rho
Usage : $factory->rho(0.3);
Function: Set the *scale* (overall evolutionary rate) of the model for the
conserved state to be the supplied number times that of the model for
the non-conserved state (default 0.3).
Returns : number (default undef)
Args : None to get, number (between 0 and 1) to set
=cut
sub rho {
my ($self, $num) = @_;
if (defined ($num)) {
($num > 0 && $num < 1) || $self->throw("rho value must be between 0 and 1, exclusive");
$self->{rho} = $num;
}
return $self->{rho} || return;
}
=head2 run
Title : run
Usage : $result = $factory->run($fasta_align_file, $newick_tree_file);
-or-
$result = $factory->run($align_object, $tree_object);
-or-
$result = $factory->run($align_object, $db_taxonomy_object);
Function: Runs phastCons on an alignment to find the most conserved regions
('footprinting').
Returns : array of Bio::SeqFeature::Annotated (one feature per alignment
sequence and prediction)
Args : The first argument represents an alignment, the second argument
a species tree.
The alignment can be provided as a multi-fasta format alignment
filename, or a Bio::Align::AlignI compliant object (eg. a
Bio::SimpleAlign).
The species tree can be provided as a newick format tree filename
or a Bio::Tree::TreeI compliant object. Alternatively a
Bio::DB::Taxonomy object can be supplied, in which case the species
tree will be generated by using the alignment sequence names as
species names and looking for those in the supplied database.
In all cases, the alignment sequence names must correspond to node
ids in the species tree. Multi-word species names should be joined
with underscores to form the sequence names, eg. Homo_sapiens
=cut
sub run {
my ($self, $aln, $tree) = @_;
($aln && $tree) || $self->throw("alignment and tree must be supplied");
my $aln_obj = $self->_alignment($aln);
$tree = $self->_tree($tree);
# if aln was a file, set the alignment id to match file name
if (-e $aln) {
my $aln_id = basename($aln);
($aln_id) = $aln_id =~ /^([^\.]+)/;
$aln_obj->id($aln_id);
}
return $self->_run;
}
sub _run {
my $self = shift;
my $exe = $self->executable || return;
# use phyloFit to generate tree model initialization (?) using species tree
# and alignment
my $pf = Bio::Tools::Run::Phylo::Phast::PhyloFit->new(-verbose => $self->verbose, -quiet => $self->quiet);
my $init_mod = $pf->run($self->_alignment, $self->_tree) || $self->throw("phyloFit failed to work as expected, is it installed?");
# cd to a temp dir
my $temp_dir = $self->tempdir;
my $cwd = Cwd->cwd();
chdir($temp_dir) || $self->throw("Couldn't change to temp dir '$temp_dir'");
my $aln_file = $self->_write_alignment;
# do training for parameter estimation
my $command = $exe.$self->_setparams($aln_file, $init_mod);
$self->debug("phastCons training command = $command\n");
system($command) && $self->throw("phastCons training call ($command) crashed: $?");
# do the final analysis
$command = $exe.$self->_setparams($aln_file);
$self->debug("phastCons command = $command\n");
system($command) && $self->throw("phastCons call ($command) crashed: $?");
# read in most_cons.bed as the result
my $bedin = Bio::FeatureIO->new(-format => 'bed', -file => 'most_cons.bed');
# cd back to orig dir
chdir($cwd) || $self->throw("Couldn't change back to working directory '$cwd'");
my @feats = ();
my $aln = $self->_alignment;
while (my $feat = $bedin->next_feature) {
$feat->source_tag('phastCons');
my $sv = Bio::Annotation::SimpleValue->new(-tagname => 'predicted', -value => 1);
$feat->annotation->add_Annotation($sv);
# $feat->type('TF_binding_site'); causes seg fault in subsequent clone()
# features are in zero-based alignment coords; make a feature for each
# alignment sequence
foreach my $seq ($aln->each_seq) {
my $clone = clone($feat);
# $clone->type('TF_binding_site'); causes massive slowdown if you later store/retrieve these features from Bio::DB::SeqFeature database
# give it the correct id
$clone->seq_id($seq->id);
# check and correct the coords (sequence may not have the feature)
my $sloc = $seq->location_from_column($feat->start + 1) || next;
my $eloc = $seq->location_from_column($feat->end + 1) || next;
$clone->start($sloc->start - 1);
$clone->end($eloc->end - 1);
push(@feats, $clone);
}
}
return @feats;
}
=head2 _setparams
Title : _setparams
Usage : Internal function, not to be called directly
Function: Creates a string of params to be used in the command string
Returns : string of params
Args : alignment file name for result production, AND filename of phyloFit
generated init.mod file to estimate trees
=cut
sub _setparams {
my ($self, $aln_file, $init_mod) = @_;
my $param_string = $self->SUPER::_setparams(-params => [keys %PARAMS],
-switches => [keys %SWITCHES],
-double_dash => 1,
-underscore_to_dash => 1);
$param_string .= ' --no-post-probs';
my $aln_id = $self->_alignment->id;
$param_string .= " --seqname $aln_id --idpref $aln_id" if $aln_id;
$param_string .= ' --refidx 0';
my $input = ' --msa-format FASTA '.$aln_file;
if ($init_mod) {
$param_string .= ' --estimate-trees mytrees '.$input.' '.$init_mod;
}
else {
$param_string .= $input.' --most-conserved most_cons.bed --score mytrees.cons.mod,mytrees.noncons.mod';
}
return $param_string;
}
1;
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