RNA polymerase II pausing can be retained or acquired during activation of genes involved in the epithelial to mesenchymal transition.

Abstract

Promoter-proximal RNA polymerase II (Pol II) pausing is implicated in the regulation of gene transcription. However, the mechanisms of pausing including its dynamics during transcriptional responses remain to be fully understood. We performed global analysis of short capped RNAs and Pol II Chromatin Immunoprecipitation sequencing in MCF-7 breast cancer cells to map Pol II pausing across the genome, and used permanganate footprinting to specifically follow pausing during transcriptional activation of several genes involved in the epithelial to mesenchymal transition (EMT). We find that the gene for EMT master regulator Snail (SNAI1), but not Slug (SNAI2), shows evidence of Pol II pausing before activation. Transcriptional activation of the paused SNAI1 gene is accompanied by a further increase in Pol II pausing signal, whereas activation of non-paused SNAI2 gene results in the acquisition of a typical pausing signature. The increase in pausing signal reflects increased transcription initiation without changes in Pol II pausing. Activation of the heat shock HSP70 gene involves pausing release that speeds up Pol II turnover, but does not change pausing location. We suggest that Pol II pausing is retained during transcriptional activation and can further undergo regulated release in a signal-specific manner.

• Authors: Ann Samarakkody, Ata Abbas, Adam Scheidegger, Jessica Warns, Oscar Nnoli, Bradley Jokinen, Kris Zarns, Brooke Kubat, Archana Dhasarathy, Sergei Nechaev
• Journal: Nucleic acids research (Nucleic Acids Res) Vol. 43 Issue 8 Pg. 3938-49 (Apr 30 2015) ISSN: 1362-4962 [Electronic] England
• PMID: 25820424 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S.)
• Copyright: © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

Chemical References

• HSP70 Heat-Shock Proteins
• SNAI1 protein, human
• SNAI2 protein, human
• Snail Family Transcription Factors
• Transcription Factors
• RNA Polymerase II

Topics

• Epithelial-Mesenchymal Transition (genetics)
• HSP70 Heat-Shock Proteins (genetics)
• Humans
• MCF-7 Cells
• Promoter Regions, Genetic
• RNA Polymerase II (metabolism)
• Snail Family Transcription Factors
• Transcription Factors (genetics)
• Transcriptional Activation