The structure and function of an RNA polymerase interaction domain in the PcrA/UvrD helicase.
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.
DNA:Protein Interactions Unit, School of Biochemistry, Biomedical Sciences Building, University of Bristol, Bristol BS8 1TD, UK.
Kelly
K
Sanders
Institute of Cancer Research, Chester Beatty Laboratories, 237 Fulham Road, London SW3 6JB, UK and Section of Structural Biology, Department of Medicine, Imperial College London, South Kensington Campus, London SW7 2AZ, UK.
Chia-Liang
CL
Lin
DNA:Protein Interactions Unit, School of Biochemistry, Biomedical Sciences Building, University of Bristol, Bristol BS8 1TD, UK.
Abigail J
AJ
Smith
Institute of Cancer Research, Chester Beatty Laboratories, 237 Fulham Road, London SW3 6JB, UK and Section of Structural Biology, Department of Medicine, Imperial College London, South Kensington Campus, London SW7 2AZ, UK.
Nora
N
Cronin
DNA:Protein Interactions Unit, School of Biochemistry, Biomedical Sciences Building, University of Bristol, Bristol BS8 1TD, UK.
Gemma
G
Fisher
Department of Biology, University of York, Wentworth Way, York YO10 5DD, UK.
Vasileios
V
Eftychidis
Department of Biology, University of York, Wentworth Way, York YO10 5DD, UK.
Peter
P
McGlynn
DNA:Protein Interactions Unit, School of Biochemistry, Biomedical Sciences Building, University of Bristol, Bristol BS8 1TD, UK.
Nigel J
NJ
Savery
Institute of Cancer Research, Chester Beatty Laboratories, 237 Fulham Road, London SW3 6JB, UK and Section of Structural Biology, Department of Medicine, Imperial College London, South Kensington Campus, London SW7 2AZ, UK.
Dale B
DB
Wigley
DNA:Protein Interactions Unit, School of Biochemistry, Biomedical Sciences Building, University of Bristol, Bristol BS8 1TD, UK.
Mark S
MS
Dillingham
D000596:Amino Acids / Q000737:chemistry; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism*; D004265:DNA Helicases / Q000737:chemistry* / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D001411:Geobacillus stearothermophilus / Q000201:enzymology; D008958:Models, Molecular; D011485:Protein Binding; D061805:Transcription Elongation, Genetic; D000071856:Tudor Domain
D000596:Amino Acids; D001426:Bacterial Proteins; C084217:pcrA protein, Bacteria; D012321:DNA-Directed RNA Polymerases; D004265:DNA Helicases