Abstract
The PcrA/UvrD helicase functions in multiple pathways that promote bacterial genome stability including the suppression of conflicts between replication and transcription and facilitating the repair of transcribed DNA. The reported ability of PcrA/UvrD to bind and backtrack RNA polymerase (1,2) might be relevant to these functions, but the structural basis for this activity is poorly understood. In this work, we define a minimal RNA polymerase interaction domain in PcrA, and report its crystal structure at 1.5 Å resolution. The domain adopts a Tudor-like fold that is similar to other RNA polymerase interaction domains, including that of the prototype transcription-repair coupling factor Mfd. Removal or mutation of the interaction domain reduces the ability of PcrA/UvrD to interact with and to remodel RNA polymerase complexes in vitro. The implications of this work for our understanding of the role of PcrA/UvrD at the interface of DNA replication, transcription and repair are discussed.
Highlights
Helicases are ubiquitous, abundant and diverse enzymes playing a wide variety of different roles in cellular nucleic acid metabolism [3]
An interesting example is provided by the UvrD helicase which has been implicated in nucleotide excision repair (NER), mismatch repair, homologous recombination and rolling circle replication mechanisms [7,8,9,10,11,12,13]
To confirm that the shorter Cterminal domain (CTD) construct retained the ability to interact with RNAP, we performed affinity pulldown experiments using the his-tagged PcrA-sCt protein as bait and nucleic acid-depleted cell extracts as prey [14]
Summary
Abundant and diverse enzymes playing a wide variety of different roles in cellular nucleic acid metabolism [3]. An interesting example is provided by the UvrD helicase ( annotated Helicase II, or PcrA in many gram positive bacteria including Bacillus subtilis) which has been implicated in nucleotide excision repair (NER), mismatch repair, homologous recombination and rolling circle replication mechanisms [7,8,9,10,11,12,13] This multi-functionality is reflected in the ability of UvrD/PcrA to interact physically and functionally with many different partner proteins including UvrB, MutL, MutS, RecA and RepC/D [13,14,15,16,17,18,19,20]. This ability of UvrD to remodel RNAP-DNA complexes might be relevant to the ability of
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
