As the basal bricks, the dynamics and arrangement of nucleosomes orchestrate the higher architecture of chromatin in a fundamental way, thereby affecting almost all nuclear biology processes. Thanks to its rather simple protocol, ATAC-seq has been rapidly adopted as a major tool for chromatin-accessible profiling at both bulk and single-cell level; however, to picture the arrangement of nucleosomes per se remains a challenge with ATAC-seq. We introduce a novel ATAC-seq analysis toolkit, named deNOPA, to predict nucleosome positions. Assessments showed that deNOPA not only outperformed state-of-the-art tools, but it is the only tool able to predict nucleosome position precisely with ultrasparse ATAC-seq data. The remarkable performance of deNOPA was fueled by the reads from short fragments, which compose nearly half of sequenced reads and are normally discarded from an ATAC-seq library. However, we found that the short fragment reads enrich information on nucleosome positions and that the linker regions were predicted by reads from both short and long fragments using Gaussian smoothing.
The current version of deNOPA was developed with python 2.7. Python 3 is not supported by now. Besides a python environment, you need also get the following pre-requirements installed:
Use the following commands to get deNOPA installed.
git clone https://gitee.com/bxxu/denopa.git
cd denopa
python setup.py install
usage: denopa [-h] -i INPUT [-o OUTPUT] [-b BUFFERSIZE] [-s CHROMSKIP] [-n NAME] [-m MAXLEN] [--proc PROC] [-p PARER] [-q QNFR] [-r]
Decoding the nucleosome positions with ATAC-seq data at single cell level
optional arguments:
-h, --help show this help message and exit
-i INPUT, --input INPUT
The input bam files. The files should be sorted. This argument could be given multiple times for multiple input files.
-o OUTPUT, --output OUTPUT
The directory where the output files will go to. It will be created if not exists (default .).
-b BUFFERSIZE, --bufferSize BUFFERSIZE
Number of reads buffered in reading the bam file (default 1000000).
-s CHROMSKIP, --chromSkip CHROMSKIP
Names of chromosomes skiped from the processing. Multiple values should be sepaated by ',' (default chrY,chrM).
-n NAME, --name NAME The name of the project (default deNOPA).
-m MAXLEN, --maxLen MAXLEN
The maximun fragment length in the input files (default 2000).
--proc PROC
Number of processors used in the analysis (default 1).
-p PARER, --pARER PARER
The p-value threshold used in determining the ATAC-seq reads enriched regions (ARERs, default 0.1).
-q QNFR, --qNFR QNFR
The q-value threshold used in determining the nucleosome free regions (NFRs, default 0.1).
-r, --removeIntermediateFiles
The intermediate files will be removed if this flag is set.
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