- important bug fix during FDR computation in the previous version (v0.07).
- now user can control the stringency level at which to search for NFRs - using the -l parameter.
- now performs normalization of read coverage (sequencing depth normalization) after removing the PCR duplicates.
- since v0.06 PARE supports one or more BAM files (replicates) as input. Previous versions of PARE (0.01 - 0.05) supports only two BAM files (replicates) as input. In practical terms, -j parameter is deprecated and -p parameter requires an argument now.
PARE is a computational method to Predict Active Regulatory Elements (enhancers and promoters). It implements a novel approach to detect Peak-Valley-Peak (PVP) pattern defined based on H3K4me1 and H3K4me3 signal at enhancers and promoters, respectively.
- Genomic regions enriched for H3K4me1 PVP pattern are predicted as active enhancers.
- Genomic regions enriched for H3K4me3 PVP pattern are predicted as active promoters.
0.08
Please cite:
Pundhir S, Bagger FO, Lauridsen FB, Rapin N, Porse BT. (2016) Peak-valley-peak pattern of histone modifications delineates active regulatory elements and their directionality. Nucleic Acids Res. [PMID: 27095194].
Generally, the user would be interested in following two scripts:
- pare: it is the main script to detect enhancers or promoters based on input BAM files (H3K4me1 - enhancers; H3K4me3 - promoters).
- bed2direction: this script is used to detect directionality of stable transcription at promoter regions, provided as input in BED format.
An example dataset and expected results are available at:
To install PARESuite, download PARESuite.tar.gz and unpack it. A directory, PARESuite will be created
tar -zxvf PARESuite.tar.gz
Now compile and create executable blockbuster
make or make all
Export environment variable 'PAREPATH' containing path to PARESuite installation directory
export PAREPATH=<path to PARESuite installation directory>
Add 'PAREPATH' to your 'PATH' environment variable
export PATH=$PATH:$PAREPATH/bin
Add 'PAREPATH' to your 'PERL5LIB' environment variable
export PERL5LIB=$PERL5LIB:$PAREPATH/share/perl/
To permanently add or update the environment variable(s), add the last three export commands in your ~/.bashrc file
We assume that the following programming platforms are installed and working: perl, R, and gcc. Besides, following packages should be installed.
Install the needed perl modules
sudo cpan Tie::IxHash Statistics::Basic
R modules are installed by entering R (type R on the cmdline) and then enter the following three commands (follow the instructions on the screen):
install.packages(c("ggplot2", "gridExtra", "optparse", "randomForest", "e1071"))
source("http://bioconductor.org/biocLite.R")
biocLite(c("DESeq"))
download samtools from http://sourceforge.net/projects/samtools/files/samtools/1.2/samtools-1.2.tar.bz2/download, go to the download location and do
tar xjf samtools-1.2.tar.bz2
cd samtools-1.2
make -j10 prefix=$HOME install
download bedtools from https://github.com/arq5x/bedtools2/releases/download/v2.23.0/bedtools-2.23.0.tar.gz, go to the download location and do
tar xzf BEDTools.v2.23
cd bedtools-2.23.0/
make -j 10
cp bin/* $HOME/bin
download featureCounts (subread) from http://sourceforge.net/projects/subread/files/subread-1.4.6-p4/, go to the download location and do
tar xzf subread-1.4.6-p4-Linux-x86_64.tar.gz
cd subread-1.4.6-p3-Linux-x86_64
cp bin/featureCounts $HOME/bin
download bedGraphToBigWig from http://hgdownload.soe.ucsc.edu/admin/exe/ for your operating system, go to the download location and do
cp bedGraphToBigWig $HOME/bin
chmod 755 $HOME/bin/bedGraphToBigWig
- download macs2 version 2.1.0 from https://github.com/taoliu/MACS/, go to the download location and install as mentioned in INSTALL.rst file
PARESuite is called with the following parameters
pare -i <BAM file(s)> [OPTIONS]
An usage example of PARESuite is shown below. As input, the method requires mapped reads in BAM format. An example dataset and expected results are available at http://servers.binf.ku.dk/pare/download/test_run/
pare -i data/h3k4me1_helas3_Rep1.bam,data/h3k4me1_helas3_Rep2.bam -o results -m hg19 -p 10 &> pare.log
As input, the method requires one or more BAM files correspondng to each replicate of H3K4me1 (enhancer prediction) or H3K4me3 (promoter prediction) ChIP-seq experiment. The name of the input file(s) should be formatted as
Input file name (replicate 1): <unique id><Rep1>.bam (example: h3k4me1_Rep1.bam)
Input file name (replicate 2): <unique id><Rep2>.bam (example: h3k4me1_Rep2.bam)
.
.
.
Input file name (replicate N): <unique id><RepN>.bam (example: h3k4me1_RepN.bam)
The chromosome identifier in the input BAM files should start with chr, for example as chrY and not like Y.
The results from the PARESuite are presented in two text files:
- RESULTS.TXT: main result file in BED format
For easy access, the html version of this file (RESULTS.HTML) is also available within the output directory
- RESULTS.UCSC: file to view the enhancer and promoter regions in UCSC browser
for more and latest information, please refer to http://spundhir.github.io/PARE/ or http://servers.binf.ku.dk/pare/
PARE: a computational method to Predict Active Regulatory Elements using histone marks
Copyright (C) 2015 Sachin Pundhir (pundhir@binf.ku.dk)
This program is free software: you can redistribute it and/or modify it under the terms of the GNU General Public License as published by the Free Software Foundation, either version 3 of the License, or (at your option) any later version.
This program is distributed in the hope that it will be useful, but WITHOUT ANY WARRANTY; without even the implied warranty of MERCHANTABILITY or FITNESS FOR A PARTICULAR PURPOSE. See the GNU General Public License for more details.
You should have received a copy of the GNU General Public License along with this program. If not, see <http://www.gnu.org/licenses/>.