DNA polymerase V activity is autoregulated by a novel intrinsic DNA-dependent ATPase.
Escherichia coli DNA polymerase V (pol V), a heterotrimeric complex composed of UmuD'2C, is marginally active. ATP and RecA play essential roles in the activation of pol V for DNA synthesis including translesion synthesis (TLS). We have established three features of the roles of ATP and RecA. (1) RecA-activated DNA polymerase V (pol V Mut), is a DNA-dependent ATPase; (2) bound ATP is required for DNA synthesis; (3) pol V Mut function is regulated by ATP, with ATP required to bind primer/template (p/t) DNA and ATP hydrolysis triggering dissociation from the DNA. Pol V Mut formed with an ATPase-deficient RecA E38K/K72R mutant hydrolyzes ATP rapidly, establishing the DNA-dependent ATPase as an intrinsic property of pol V Mut distinct from the ATP hydrolytic activity of RecA when bound to single-stranded (ss)DNA as a nucleoprotein filament (RecA*). No similar ATPase activity or autoregulatory mechanism has previously been found for a DNA polymerase.DOI: http://dx.doi.org/10.7554/eLife.02384.001.
Department of Biological Sciences, University of Southern California, Los Angeles, United States.
Aysen L
AL
Erdem
Department of Biological Sciences, University of Southern California, Los Angeles, United States.
Malgorzata
M
Jaszczur
Department of Biological Sciences, University of Southern California, Los Angeles, United States.
Jeffrey G
JG
Bertram
Laboratory of Genomic Integrity, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, United States.
Roger
R
Woodgate
Department of Biochemistry, University of Wisconsin-Madison, Madison, United States.
Michael M
MM
Cox
Department of Biological Sciences, University of Southern California, Los Angeles, United States Department of Chemistry, University of Southern California, Los Angeles, United States mgoodman@usc.edu.
Myron F
MF
Goodman
D000251:Adenosine Triphosphatases / Q000235:genetics* / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D006868:Hydrolysis; D009154:Mutation; D009698:Nucleoproteins / Q000235:genetics / Q000378:metabolism; D011938:Rec A Recombinases / Q000235:genetics / Q000378:metabolism
D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D009698:Nucleoproteins; D011938:Rec A Recombinases; C120926:DNA polymerase V, E coli; D004259:DNA-Directed DNA Polymerase; D000251:Adenosine Triphosphatases
ATPase; DNA polymerase V; RecA; pol V Mut