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#!/usr/bin/perl
################################################################
# copyright (c) 2010, 2014 by William R. Pearson and The Rector &
# Visitors of the University of Virginia */
################################################################
# Licensed under the Apache License, Version 2.0 (the "License");
# you may not use this file except in compliance with the License.
# You may obtain a copy of the License at
#
# http://www.apache.org/licenses/LICENSE-2.0
#
# Unless required by applicable law or agreed to in writing,
# software distributed under this License is distributed on an "AS
# IS" BASIS, WITHOUT WRRANTIES OR CONDITIONS OF ANY KIND, either
# express or implied. See the License for the specific language
# governing permissions and limitations under the License.
################################################################
## usage - expand_refseq_isoforms.pl [--prim_acc] up_hits.file > refseq_isoforms.file
##
## take a fasta36 -e expand.sh result file of the form:
## NP_000552|<tab>1.1e-50
##
## and extract the accession number, looking it up from the an SQL
## table $table -- in this case "annot2_iso" to provide Uniprot
## isoforms based on a uniprot accession.
##
## if --prim_acc, then the primary accession (used to find the isoforms) is added to the isoform seq_id, e.g.
## NP_000552 has isoforms: with --prim_acc the identifiers become:
## >iso|XP_005270839 >iso|NP_666533|NP_666533::NP_000552
## >iso|NP_666533 >iso|NP_666533|NP_666533::NP_000552
##
use warnings;
use strict;
use Getopt::Long;
use Pod::Usage;
use DBI;
my ($host, $db, $port, $user, $pass) = ("xdb", "seqdb_demo2", 0, "web_user", "fasta_www");
$host = 'wrpxdb.its.virginia.edu';
my ($help, $shelp) = (0,0);
my ($e_thresh, $prim_acc) = (1e-6, 0);
GetOptions(
"h" => \$shelp,
"help" => \$help,
"host=s" => \$host,
"prim_acc!" => \$prim_acc,
"db=s" => \$db,
"expect|evalue|e_thresh=f" => \$e_thresh,
"user=s" => \$user,
"password=s" => \$pass,
"port=i" => \$port,
);
pod2usage(1) if $shelp;
pod2usage(exitstatus => 0, verbose => 2) if $help;
pod2usage(1) unless (@ARGV || -p STDIN || -f STDIN);
my $dbh = DBI->connect("dbi:mysql:host=$host:$db",
$user, $pass,
{ RaiseError => 1, AutoCommit => 1}
) or die $DBI::errstr;
my %sth = (
seed2link_acc => "SELECT distinct(r1.acc) FROM refseq_NP_NM_geneid as r1 join refseq_NP_NM_geneid as r2 using(gene_id) WHERE r2.acc=?",
link2seq => "SELECT db, acc, descr, seq FROM annot JOIN protein USING(prot_id) WHERE acc=?"
);
for my $sth (keys(%sth)) {
$sth{$sth} = $dbh->prepare($sth{$sth});
}
my %acc_uniq = ();
# get the query
my ($query, $eval_arg) = @ARGV;
$eval_arg = 1e-10 unless $eval_arg;
$query =~ s/^>// if ($query);
my @link_lines = ();
#if it's a file I can open, read and parse it
unless ($query && ($query =~ m/[\|:]/ ||
$query =~ m/^[NXW]P_/ ||
$query =~ m/^[OPQ][0-9][A-Z0-9]{3}[0-9]|[A-NR-Z][0-9]([A-Z][A-Z0-9]{2}[0-9]){1,2}\s/)) {
while (my $a_line = <>) {
$a_line =~ s/^>//;
chomp $a_line;
push @link_lines, $a_line;
}
}
else {
push @link_lines, "$query\t$eval_arg";
}
for my $line ( @link_lines ) {
my ($hit, $e_val) = split(/\t/,$line);
if ($e_val <= $e_thresh) {
process_line($hit,$sth{seed2link_acc});
}
}
my $have_isoforms = 0;
for my $acc ( keys %acc_uniq ) {
$sth{link2seq}->execute($acc);
while (my $row_href = $sth{link2seq}->fetchrow_hashref ) {
my $id_str = $acc;
if ($prim_acc) {
$id_str .= "::".$acc_uniq{$acc};
printf(">%s|%s|%s %s\n","iso",$acc,$id_str,$row_href->{descr});
}
else {
printf(">%s|%s %s\n","iso",$acc,$row_href->{descr});
}
my $iso_seq = $row_href->{seq};
$iso_seq =~ s/(.{60})/$1\n/g;
print "$iso_seq\n";
}
$sth{link2seq}->finish();
}
$dbh->disconnect();
exit(4) unless $have_isoforms;
sub process_line{
my ($seqid,$sth_acc)=@_;
my $sth = $sth_acc;
my ($db, $link_acc, $link_id) = ("","","");
if ($seqid =~ m/\|/) {
($db, $link_acc) = split('\|',$seqid);
$link_acc =~ s/\.\d+$//;
$sth_acc->execute($link_acc);
}
else {
$link_acc = $seqid;
$link_acc =~ s/\.\d+$//;
$sth_acc->execute($link_acc);
}
while (my ($acc) = $sth->fetchrow_array()) {
next if ($acc eq $link_acc);
$acc_uniq{$acc} = $link_acc unless $acc_uniq{$acc};
}
$sth->finish();
}
__END__
=pod
=head1 NAME
expand_refseq_isoforms.pl expand_file.tab
=head1 SYNOPSIS
expand_refseq_isoforms.pl expand_file.tab
=head1 OPTIONS
-h short help
--help include description
--evalue E()-value threshold for expansion
--prim_acc : show primary accession as part of sequence identifier
>iso|XP_005270839|XP_005270839::NP_000552 becomes >iso|XP_005270839|XP_005270839::NP_000552
--host, --user, --password, --port --db : info for mysql database
=head1 DESCRIPTION
C<expand_refseq_isoforms.pl> uses protein isoform tables in an SQL database to identify and extract
isoforms of proteins in a reference protein sequence database.
C<expand_refseq_isoforms.pl> takes a file with sequece identifiers and E()-values of the form:
ref|NP_000552 1e-10
NP_000662
Lines with E()-values less than --evalue (1E-6 by default) are used to
identify protein isoforms, which are included in the set of sequences to be aligned.
C<expand_refseq_isoforms.pl> is designed to be used by the B<FASTA> programs with
the C<-e expand_refseq_isoforms.pl> option.
=head1 AUTHOR
William R. Pearson, wrp@virginia.edu
=cut
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