The conserved C-terminus of the PcrA/UvrD helicase interacts directly with RNA polymerase

Emma J Gwynn; Abigail J Smith; Colin P Guy; Nigel J Savery; Peter McGlynn; Mark S Dillingham

doi:10.1371/journal.pone.0078141

The conserved C-terminus of the PcrA/UvrD helicase interacts directly with RNA polymerase

Emma J Gwynn, Abigail J Smith, Colin P Guy, Nigel J Savery, Peter McGlynn, Mark S Dillingham

Medical Sciences

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Abstract

UvrD-like helicases play diverse roles in DNA replication, repair and recombination pathways. An emerging body of evidence suggests that their different cellular functions are directed by interactions with partner proteins that target unwinding activity to appropriate substrates. Recent studies in E. coli have shown that UvrD can act as an accessory replicative helicase that resolves conflicts between the replisome and transcription complexes, but the mechanism is not understood. Here we show that the UvrD homologue PcrA interacts physically with B. subtilis RNA polymerase, and that an equivalent interaction is conserved in E. coli where UvrD, but not the closely related helicase Rep, also interacts with RNA polymerase. The PcrA-RNAP interaction is direct and independent of nucleic acids or additional mediator proteins. A disordered but highly conserved C-terminal region of PcrA, which distinguishes PcrA/UvrD from otherwise related enzymes such as Rep, is both necessary and sufficient for interaction with RNA polymerase.

Original language	English
Article number	e78141
Number of pages	11
Journal	PloS ONE
Volume	8
Issue number	10
DOIs	https://doi.org/10.1371/journal.pone.0078141
Publication status	Published - 16 Oct 2013

Bibliographical note

Copyright: © 2013 Gwynn et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Funding: This work was supported by a Wellcome Trust project grant to MD (Reference: 077368), an ERC starting grant to MD (Acronym: SM-DNA-REPAIR) and a BBSRC project grant to PM, NS and MD (Reference: BB/I003142/1). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Keywords

couple DNA-repair
single-stranded-dna
stearothermophilus pcra helicase
Escherichia-coli
bacillus-subtilis
UVRD helicase
crystal-structures
structural basis
mismatch repair
in-vitro

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10.1371/journal.pone.0078141

The conserved c-terminus
This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. http://creativecommons.org/licenses/by/3.0/
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abstract = "UvrD-like helicases play diverse roles in DNA replication, repair and recombination pathways. An emerging body of evidence suggests that their different cellular functions are directed by interactions with partner proteins that target unwinding activity to appropriate substrates. Recent studies in E. coli have shown that UvrD can act as an accessory replicative helicase that resolves conflicts between the replisome and transcription complexes, but the mechanism is not understood. Here we show that the UvrD homologue PcrA interacts physically with B. subtilis RNA polymerase, and that an equivalent interaction is conserved in E. coli where UvrD, but not the closely related helicase Rep, also interacts with RNA polymerase. The PcrA-RNAP interaction is direct and independent of nucleic acids or additional mediator proteins. A disordered but highly conserved C-terminal region of PcrA, which distinguishes PcrA/UvrD from otherwise related enzymes such as Rep, is both necessary and sufficient for interaction with RNA polymerase.",

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KW - structural basis

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KW - in-vitro

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The conserved C-terminus of the PcrA/UvrD helicase interacts directly with RNA polymerase

Abstract

Bibliographical note

Keywords

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