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Journal of Bacteriology, March 2005, p. 1974-1984, Vol. 187, No. 6
0021-9193/05/$08.00+0     doi:10.1128/JB.187.6.1974-1984.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

In Vivo Evidence for a recA-Independent Recombination Process in Escherichia coli That Permits Completion of Replication of DNA Containing UV Damage in Both Strands

Ali I. Ozgenc, Edward S. Szekeres,{dagger} and Christopher W. Lawrence{ddagger}*

Department of Biochemistry & Biophysics, University of Rochester School of Medicine & Dentistry, Rochester, New York

Received 21 October 2004/ Accepted 13 December 2004

We have investigated recombination mechanisms promoting the completion of replication in the face of unrepaired DNA damage by transforming an isogenic set of uvrA6 excision-defective Escherichia coli strains with pUC-based plasmids in which each strand carried, at staggered positions, a single thymine-thymine pyrimidine (6-4) pyrimidinone lesion. The distance between the lesions was 28 or 8 bp in one orientation relative to the unidirectional ColE1 origin of replication or, in the other orientation, 30 or 10 bp. C-C mismatches placed opposite each of the T-T photoproducts permit unambiguous detection of the three events that can lead to the completion of replication: sister-strand recombination, translesion replication (TR) on the leading strand, and TR on the lagging strand. We find that E. coli possesses a largely constitutive, recA-independent sister-strand recombination mechanism that allows 9% or more of these severely compromised plasmids to be fully replicated. In one orientation, such recombination depends partly on recG and priA but not on ruvA, ruvB, ruvC, or mutS and is largely independent of recF. In the other orientation, recombination is dependent on none of the genes. The strains used did not contain the cryptic phage encoding recET, which encodes enzymes that promote interplasmid recombination. The nature of the recA-independent recombination mechanism is not known but could perhaps result from a template-strand-switching, or copy choice, process.

* Corresponding author. Mailing address: Department of Biochemistry and Biophysics, University of Rochester Medical Center, 601 Elmwood Ave., Rochester, NY 14642. Phone: (585) 275-2948. Fax: (585) 275-6007. E-mail: christopher_lawrence{at}urmc.rochester.edu.

{dagger} Present address: Department of Pathology, University of Washington, Seattle, WA 98195.

{ddagger} Present address: CuraGen Corporation, Branford, CT 06405.

Journal of Bacteriology, March 2005, p. 1974-1984, Vol. 187, No. 6
0021-9193/05/$08.00+0     doi:10.1128/JB.187.6.1974-1984.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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