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2014 ; 15 Suppl 1
(Suppl 1
): S1
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An improved ChIP-seq peak detection system for simultaneously identifying
post-translational modified transcription factors by combinatorial fusion, using
SUMOylation as an example
#MMPMID24564277
Cheng CY
; Chu CH
; Hsu HW
; Hsu FR
; Tang CY
; Wang WC
; Kung HJ
; Chang PC
BMC Genomics
2014[]; 15 Suppl 1
(Suppl 1
): S1
PMID24564277
show ga
BACKGROUND: Post-translational modification (PTM) of transcriptional factors and
chromatin remodelling proteins is recognized as a major mechanism by which
transcriptional regulation occurs. Chromatin immunoprecipitation (ChIP) in
combination with high-throughput sequencing (ChIP-seq) is being applied as a gold
standard when studying the genome-wide binding sites of transcription factor
(TFs). This has greatly improved our understanding of protein-DNA interactions on
a genomic-wide scale. However, current ChIP-seq peak calling tools are not
sufficiently sensitive and are unable to simultaneously identify
post-translational modified TFs based on ChIP-seq analysis; this is largely due
to the wide-spread presence of multiple modified TFs. Using SUMO-1 modification
as an example; we describe here an improved approach that allows the simultaneous
identification of the particular genomic binding regions of all TFs with SUMO-1
modification. RESULTS: Traditional peak calling methods are inadequate when
identifying multiple TF binding sites that involve long genomic regions and
therefore we designed a ChIP-seq processing pipeline for the detection of peaks
via a combinatorial fusion method. Then, we annotate the peaks with known
transcription factor binding sites (TFBS) using the Transfac Matrix Database
(v7.0), which predicts potential SUMOylated TFs. Next, the peak calling result
was further analyzed based on the promoter proximity, TFBS annotation, a
literature review, and was validated by ChIP-real-time quantitative PCR (qPCR)
and ChIP-reChIP real-time qPCR. The results show clearly that SUMOylated TFs are
able to be pinpointed using our pipeline. CONCLUSION: A methodology is presented
that analyzes SUMO-1 ChIP-seq patterns and predicts related TFs. Our analysis
uses three peak calling tools. The fusion of these different tools increases the
precision of the peak calling results. TFBS annotation method is able to predict
potential SUMOylated TFs. Here, we offer a new approach that enhances ChIP-seq
data analysis and allows the identification of multiple SUMOylated TF binding
sites simultaneously, which can then be utilized for other functional PTM binding
site prediction in future.