Genetics, Vol 143, 5-13, Copyright © 1996

INVESTIGATIONS

Differential Suppression of priA2::kan Phenotypes in Escherichia coli K-12 by Mutations in priA, lexA, and dnaC

S. J. Sandler, H. S. Samra and A. J. Clark
Department of Molecular and Cell Biology, Division of Genetics, University of California, Berkeley, California 94720-3202

First identified as an essential component of the {phi}X174 in vitro DNA replication system, PriA has ATPase, helicase, translocase, and primosome-assembly activities. priA1::kan strains of Escherichia coli are sensitive to UV irradiation, deficient in homologous recombination following transduction, and filamentous. priA2::kan strains have eightfold higher levels of uninduced SOS expression than wild type. We show that (1) priA1::kan strains have eightfold higher levels of uninduced SOS expression, (2) priA2::kan strains are UV(S) and Rec(-), (3) lexA3 suppresses the high basal levels of SOS expression of a priA2::kan strain, and (4) plasmid-encoded priA300 (K230R), a mutant allele retaining only the primosome-assembly activity of priA(+), restores both UV(R) and Rec(+) phenotypes to a priA2::kan strain. Finally, we have isolated 17 independent UV(R) Rec(+) revertants of priA2::kan strains that carry extragenic suppressors. All 17 map in the C-terminal half of the dnaC gene. DnaC loads the DnaB helicase onto DNA as a prelude for primosome assembly and DNA replication. We conclude that priA's primosome-assembly activity is essential for DNA repair and recombination and that the dnaC suppressor mutations allow these processes to occur in the absence of priA.

This article has been cited by other articles:

				Q. Shi, A. R. Parks, B. D. Potter, I. J. Safir, Y. Luo, B. M. Forster, and J. E. Peters DNA Damage Differentially Activates Regional Chromosomal Loci for Tn7 Transposition in Escherichia coli Genetics, July 1, 2008; 179(3): 1237 - 1250. [Abstract] [Full Text] [PDF]

				I. Ivancic-Bace, I. Vlasic, G. Cogelja-Cajo, K. Brcic-Kostic, and E. Salaj-Smic Roles of PriA Protein and Double-Strand DNA Break Repair Functions in UV-Induced Restriction Alleviation in Escherichia coli Genetics, December 1, 2006; 174(4): 2137 - 2149. [Abstract] [Full Text] [PDF]

				C. J. Cadman, M. Lopper, P. B. Moon, J. L. Keck, and P. McGlynn PriB Stimulates PriA Helicase via an Interaction with Single-stranded DNA J. Biol. Chem., December 2, 2005; 280(48): 39693 - 39700. [Abstract] [Full Text] [PDF]

				Z. Pang, R. Chen, D. Manna, and N. P. Higgins A Gyrase Mutant with Low Activity Disrupts Supercoiling at the Replication Terminus J. Bacteriol., November 15, 2005; 187(22): 7773 - 7783. [Abstract] [Full Text] [PDF]

				K. A. Kline and H. S. Seifert Mutation of the priA Gene of Neisseria gonorrhoeae Affects DNA Transformation and DNA Repair J. Bacteriol., August 1, 2005; 187(15): 5347 - 5355. [Abstract] [Full Text] [PDF]

				K. A. Kline and H. S. Seifert Role of the Rep Helicase Gene in Homologous Recombination in Neisseria gonorrhoeae J. Bacteriol., April 15, 2005; 187(8): 2903 - 2907. [Abstract] [Full Text] [PDF]

				S. J. Sandler Requirements for Replication Restart Proteins During Constitutive Stable DNA Replication in Escherichia coli K-12 Genetics, April 1, 2005; 169(4): 1799 - 1806. [Abstract] [Full Text] [PDF]

				T. Shibata, T. Hishida, Y. Kubota, Y.-W. Han, H. Iwasaki, and H. Shinagawa Functional overlap between RecA and MgsA (RarA) in the rescue of stalled replication forks in Escherichia coli Genes Cells, March 1, 2005; 10(3): 181 - 191. [Abstract] [Full Text] [PDF]

				C. J. Cadman and P. McGlynn PriA helicase and SSB interact physically and functionally Nucleic Acids Res., December 2, 2004; 32(21): 6378 - 6387. [Abstract] [Full Text] [PDF]

				B. Michel, G. Grompone, M.-J. Flores, and V. Bidnenko Multiple pathways process stalled replication forks PNAS, August 31, 2004; 101(35): 12783 - 12788. [Abstract] [Full Text] [PDF]

				J. D. McCool, C. C. Ford, and S. J. Sandler A dnaT Mutant With Phenotypes Similar to Those of a priA2::kan Mutant in Escherichia coli K-12 Genetics, June 1, 2004; 167(2): 569 - 578. [Abstract] [Full Text] [PDF]

				R. Harinarayanan and J. Gowrishankar A dnaC Mutation in Escherichia coli That Affects Copy Number of ColE1-Like Plasmids and the PriA-PriB (but Not Rep-PriC) Pathway of Chromosomal Replication Restart Genetics, March 1, 2004; 166(3): 1165 - 1176. [Abstract] [Full Text] [PDF]

				G. Grompone, V. Bidnenko, S. D. Ehrlich, and B. Michel PriA Is Essential for Viability of the Escherichia coli Topoisomerase IV parE10(Ts) Mutant J. Bacteriol., February 15, 2004; 186(4): 1197 - 1199. [Abstract] [Full Text] [PDF]

				T. Mizukoshi, T. Tanaka, K.-i. Arai, D. Kohda, and H. Masai A Critical Role of the 3' Terminus of Nascent DNA Chains in Recognition of Stalled Replication Forks J. Biol. Chem., October 24, 2003; 278(43): 42234 - 42239. [Abstract] [Full Text] [PDF]

				T. Tanaka, T. Mizukoshi, C. Taniyama, D. Kohda, K.-i. Arai, and H. Masai DNA Binding of PriA Protein Requires Cooperation of the N-terminal D-loop/Arrested-fork Binding and C-terminal Helicase Domains J. Biol. Chem., October 4, 2002; 277(41): 38062 - 38071. [Abstract] [Full Text] [PDF]

				P. Polard, S. Marsin, S. McGovern, M. Velten, D. B. Wigley, S. D. Ehrlich, and C. Bruand Restart of DNA replication in Gram-positive bacteria: functional characterisation of the Bacillussubtilis PriA initiator Nucleic Acids Res., April 1, 2002; 30(7): 1593 - 1605. [Abstract] [Full Text] [PDF]

				D. A. Cano, M. G. Pucciarelli, F. Garcia-del Portillo, and J. Casadesus Role of the RecBCD Recombination Pathway in Salmonella Virulence J. Bacteriol., January 15, 2002; 184(2): 592 - 595. [Abstract] [Full Text] [PDF]

				S. Marsin, S. McGovern, S. D. Ehrlich, C. Bruand, and P. Polard Early Steps of Bacillus subtilis Primosome Assembly J. Biol. Chem., November 30, 2001; 276(49): 45818 - 45825. [Abstract] [Full Text] [PDF]

				B. Michel, M.-J. Flores, E. Viguera, G. Grompone, M. Seigneur, and V. Bidnenko Rescue of arrested replication forks by homologous recombination PNAS, July 17, 2001; 98(15): 8181 - 8188. [Abstract] [Full Text] [PDF]

				J. D. McCool and S. J. Sandler Effects of mutations involving cell division, recombination, and chromosome dimer resolution on a priA2::kan mutant PNAS, July 17, 2001; 98(15): 8203 - 8210. [Abstract] [Full Text] [PDF]

				S. A. Sciochetti, G. W. Blakely, and P. J. Piggot Growth Phase Variation in Cell and Nucleoid Morphology in a Bacillus subtilis recA Mutant J. Bacteriol., May 1, 2001; 183(9): 2963 - 2968. [Abstract] [Full Text]

				L. SaiSree, M. Reddy, and J. Gowrishankar lon Incompatibility Associated with Mutations Causing SOS Induction: Null uvrD Alleles Induce an SOS Response in Escherichia coli J. Bacteriol., June 1, 2000; 182(11): 3151 - 3157. [Abstract] [Full Text]

				S. J. Sandler Multiple Genetic Pathways for Restarting DNA Replication Forks in Escherichia coli K-12 Genetics, June 1, 2000; 155(2): 487 - 497. [Abstract] [Full Text]

				L. Xu and K. J. Marians Purification and Characterization of DnaC810, a Primosomal Protein Capable of Bypassing PriA Function J. Biol. Chem., March 10, 2000; 275(11): 8196 - 8205. [Abstract] [Full Text] [PDF]

				S. J. Sandler and K. J. Marians Role of PriA in Replication Fork Reactivation in Escherichia coli J. Bacteriol., January 1, 2000; 182(1): 9 - 13. [Full Text]

				R. Rothstein, B. Michel, and S. Gangloff Replication fork pausing and recombination or "gimme a break" Genes & Dev., January 1, 2000; 14(1): 1 - 10. [Full Text]

				A. Kuzminov Recombinational Repair of DNA Damage in Escherichia coli and Bacteriophage lambda Microbiol. Mol. Biol. Rev., December 1, 1999; 63(4): 751 - 813. [Abstract] [Full Text] [PDF]

				A. R. Poteete, A. C. Fenton, and K. C. Murphy Roles of RuvC and RecG in Phage lambda Red-Mediated Recombination J. Bacteriol., September 1, 1999; 181(17): 5402 - 5408. [Abstract] [Full Text]

				P. Nurse, J. Liu, and K. J. Marians Two Modes of PriA Binding to DNA J. Biol. Chem., August 27, 1999; 274(35): 25026 - 25032. [Abstract] [Full Text] [PDF]

				J. Liu and K. J. Marians PriA-directed Assembly of a Primosome on D Loop DNA J. Biol. Chem., August 27, 1999; 274(35): 25033 - 25041. [Abstract] [Full Text] [PDF]

				P. L. Foster and W. A. Rosche Increased Episomal Replication Accounts for the High Rate of Adaptive Mutation in recD Mutants of Escherichia coli Genetics, May 1, 1999; 152(1): 15 - 30. [Abstract] [Full Text]

				J. Liu, L. Xu, S. J. Sandler, and K. J. Marians Replication fork assembly at recombination intermediates is required for bacterial growth PNAS, March 30, 1999; 96(7): 3552 - 3555. [Abstract] [Full Text] [PDF]

				J. Courcelle, D. J. Crowley, and P. C. Hanawalt Recovery of DNA Replication in UV-Irradiated Escherichia coli Requires both Excision Repair and RecF Protein Function J. Bacteriol., February 1, 1999; 181(3): 916 - 922. [Abstract] [Full Text]

				A. Kuzminov and F. W. Stahl Double-strand end repair via the RecBC pathway in Escherichia coli primes DNA replication Genes & Dev., February 1, 1999; 13(3): 345 - 356. [Abstract] [Full Text]

				S. Stambuk and M. Radman Mechanism and Control of Interspecies Recombination in Escherichia coli. I. Mismatch Repair, Methylation, Recombination and Replication Functions Genetics, October 1, 1998; 150(2): 533 - 542. [Abstract] [Full Text]

				C. H. Gross and S. Shuman The Nucleoside Triphosphatase and Helicase Activities of Vaccinia Virus NPH-II Are Essential for Virus Replication J. Virol., June 1, 1998; 72(6): 4729 - 4736. [Abstract] [Full Text] [PDF]

				J. Y. Ng and K. J. Marians The Ordered Assembly of the phi X174-type Primosome. I. ISOLATION AND IDENTIFICATION OF INTERMEDIATE PROTEIN-DNA COMPLEXES J. Biol. Chem., June 28, 1996; 271(26): 15642 - 15648. [Abstract] [Full Text] [PDF]

				J. Y. Ng and K. J. Marians The Ordered Assembly of the phi X174-type Primosome. II. PRESERVATION OF PRIMOSOME COMPOSITION FROM ASSEMBLY THROUGH REPLICATION J. Biol. Chem., June 28, 1996; 271(26): 15649 - 15655. [Abstract] [Full Text] [PDF]