Alternative DNA secondary structure formation affects RNA polymerase II promoter-proximal pausing in human

Genome Biology
Karol SzlachtaYuh-Hwa Wang

Abstract

Alternative DNA secondary structures can arise from single-stranded DNA when duplex DNA is unwound during DNA processes such as transcription, resulting in the regulation or perturbation of these processes. We identify sites of high propensity to form stable DNA secondary structure across the human genome using Mfold and ViennaRNA programs with parameters for analyzing DNA. The promoter-proximal regions of genes with paused transcription are significantly and energetically more favorable to form DNA secondary structure than non-paused genes or genes without RNA polymerase II (Pol II) binding. Using Pol II ChIP-seq, GRO-seq, NET-seq, and mNET-seq data, we arrive at a robust set of criteria for Pol II pausing, independent of annotation, and find that a highly stable secondary structure is likely to form about 10-50 nucleotides upstream of a Pol II pausing site. Structure probing data confirm the existence of DNA secondary structures enriched at the promoter-proximal regions of paused genes in human cells. Using an in vitro transcription assay, we demonstrate that Pol II pausing at HSPA1B, a human heat shock gene, is affected by manipulating DNA secondary structure upstream of the pausing site. Our results indicate alternative DNA...Continue Reading

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Citations

Jan 27, 2019·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Zhi-Yin SunTian-Miao Ou
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Datasets Mentioned

BETA
GM12878

Methods Mentioned

BETA
in vitro transcription
ChIP-seq
ChIP
immunoprecipitation
footprinting
PCR

Software Mentioned

BEDTtools
Mfold
pyplot
ViennaRNA package
quadrulex
RNAfold
UCSC Genome Browser
ViennaRNA
mNET
quadparser

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