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RecA acts in trans to allow replication of damaged DNA by DNA polymerase V

Nature volume 442, pages 883–887 (2006)Cite this article

Abstract

The DNA polymerase V (pol V) and RecA proteins are essential components of a mutagenic translesion synthesis pathway in Escherichia coli designed to cope with DNA damage. Previously, it has been assumed that RecA binds to the DNA template strand being copied. Here we show, however, that pol-V-catalysed translesion synthesis, in the presence or absence of the β-processivity-clamp, occurs only when RecA nucleoprotein filaments assemble or RecA protomers bind on separate single-stranded (ss)DNA molecules in trans. A 3′-proximal RecA filament end on trans DNA is essential for stimulation; however, synthesis is strengthened by further pol V–RecA interactions occurring elsewhere along a trans nucleoprotein filament. We suggest that trans-stimulation of pol V by RecA bound to ssDNA reflects a distinctive regulatory mechanism of mutation that resolves the paradox of RecA filaments assembled in cis on a damaged template strand obstructing translesion DNA synthesis despite the absolute requirement of RecA for SOS mutagenesis.

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Figure 1: RecA–ssDNA transactivation of pol V on damaged and undamaged DNA templates.
Figure 2: Kinetics of pol V transactivation by RecA–ssDNA.
Figure 3: A RecA filament end oriented 3′ on trans DNA is required for efficient stimulation of pol V.
Figure 4: Models depicting RecA–DNA transactivation of pol-V-catalysed translesion DNA synthesis.

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Acknowledgements

This work was supported by National Institutes of Health grants to M.F.G. and M.M.C., and funds from the NICHD/NIH Intramural Research Program to R.W.

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Authors and Affiliations

  1. Department of Biological Sciences and Chemistry, University of Southern California, University Park, California, 90089-2910, Los Angeles, USA

    Katharina Schlacher & Myron F. Goodman

  2. Department of Biochemistry, University of Wisconsin-Madison, Wisconsin, 53706, Madison, USA

    Michael M. Cox

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

    Roger Woodgate

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  1. Katharina Schlacher
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Correspondence to Myron F. Goodman.

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Supplementary information

Supplementary Figures

This file contains a background figure (Supplementary Figure 1), supportive data (Supplementary Figures 2–12) and DNA sequences (Figure 13). (PDF 1094 kb)

Supplementary Methods

This file contains additional details of the Methods used in this study. (PDF 56 kb)

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Schlacher, K., Cox, M., Woodgate, R. et al. RecA acts in trans to allow replication of damaged DNA by DNA polymerase V. Nature 442, 883–887 (2006). https://doi.org/10.1038/nature05042

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