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A model for SOS-lesion-targeted mutations in Escherichia coli

Nature volume 409, pages 366–370 (2001)Cite this article

Abstract

The UmuD′2C protein complex (Escherichia coli pol V)1,2,3 is a low-fidelity DNA polymerase (pol) that copies damaged DNA in the presence of RecA, single-stranded-DNA binding protein (SSB) and the β,γ-processivity complex of E. coli pol III (ref. 4). Here we propose a model to explain SOS-lesion-targeted mutagenesis, assigning specific biochemical functions for each protein during translesion synthesis. (SOS lesion-targeted mutagenesis occurs when pol V is induced as part of the SOS response to DNA damage and incorrectly incorporates nucleotides opposite template lesions.) Pol V plus SSB catalyses RecA filament disassembly in the 3′ to 5′ direction on the template, ahead of the polymerase, in a reaction that does not involve ATP hydrolysis. Concurrent ATP-hydrolysis-driven filament disassembly in the 5′ to 3′ direction results in a bidirectional stripping of RecA from the template strand. The bidirectional collapse of the RecA filament restricts DNA synthesis by pol V to template sites that are proximal to the lesion, thereby minimizing the occurrence of untargeted mutations at undamaged template sites.

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Figure 1: Effect of RecA, SSB and β,γ-complex on replication of lesion-containing and natural primer/template (p/t) DNA.
Figure 2: Dependence of pol V-catalysed TLS and total DNA synthesis on RecA, SSB and β,γ-complex. a, RecA protein concentration was varied while maintaining constant concentrations of SSB (400 nM), β- (400 nM) and γ-complex (100 nM).
Figure 3: Nuclease protection analysis of pol V plus SSB 3′ to 5′ disassembly of a stabilized RecA filament.
Figure 4: Cowcatcher model for TLS by the pol V mutasome.

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Acknowledgements

This work was supported by National Institutes of Health grants to M.F.G. and M.O. P.P. was supported on an NIH-NIA postdoctoral training grant, and J.G.B. was supported on a National Institute of Dental and Craniofacial Research predoctoral training grant.

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Authors and Affiliations

  1. Department of Biological Sciences and Chemistry, Hedco Molecular Biology Laboratories, University of Southern California, University Park, Los Angeles, 90089-1340, California, USA

    Phuong Pham, Jeffrey G. Bertram & Myron F. Goodman

  2. Rockefeller University and Howard Hughes Medical Institute, New York, 10021, New York, USA

    Mike O'Donnell

  3. Section on DNA Replication, Repair and Mutagenesis, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, 20892-2725, Maryland, USA

    Roger Woodgate

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  1. Phuong Pham
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  5. Myron F. Goodman
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Correspondence to Myron F. Goodman.

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Pham, P., Bertram, J., O'Donnell, M. et al. A model for SOS-lesion-targeted mutations in Escherichia coli. Nature 409, 366–370 (2001). https://doi.org/10.1038/35053116

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