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Roles of E. coli DNA polymerases IV and V in lesion-targeted and untargeted SOS mutagenesis

Nature volume 404, pages 1014–1018 (2000)Cite this article

Abstract

The expression of the Escherichia coli DNA polymerases pol V (UmuD′2C complex)1,2 and pol IV (DinB)3 increases in response to DNA damage4. The induction of pol V is accompanied by a substantial increase in mutations targeted at DNA template lesions in a process called SOS-induced error-prone repair4. Here we show that the common DNA template lesions, TT (6–4) photoproducts, TT cis–syn photodimers and abasic sites, are efficiently bypassed within 30 seconds by pol V in the presence of activated RecA protein (RecA*), single-stranded binding protein (SSB) and pol III's processivity β,γ-complex. There is no detectable bypass by either pol IV or pol III on this time scale. A mutagenic ‘signature’ for pol V is its incorporation of guanine opposite the 3′-thymine of a TT (6–4) photoproduct, in agreement with mutational spectra. In contrast, pol III and pol IV incorporate adenine almost exclusively. When copying undamaged DNA, pol V exhibits low fidelity with error rates of around 10-3 to 10-4, with pol IV being 5- to 10-fold more accurate. The effects of RecA protein on pol V, and β,γ-complex on pol IV, cause a 15,000- and 3,000-fold increase in DNA synthesis efficiency, respectively. However, both polymerases exhibit low processivity, adding 6 to 8 nucleotides before dissociating. Lesion bypass by pol V does not require β,γ-complex in the presence of non-hydrolysable ATPγS, indicating that an intact RecA filament may be required for translesion synthesis.

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Figure 1: Comparison of translesion synthesis by pol III HE, pol IV and pol V Mut.
Figure 2: Incorporation of A compared with G opposite a TT (6–4) photoproduct.
Figure 3: Analysis of the processivity of pol IV and pol V with or without SSB and β,γ-complex.
Figure 4: Effect of RecA and β,γ-complex on incorporation of correct dAMP at an undamaged template by pol V and pol IV.
Figure 5: Role of β,γ-complex in abasic site bypass by pol V in the presence of ATP or ATP-γS.

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Acknowledgements

This work was supported by National Institutes of Health grants to M.F.G., M.O. and J.-S.T.).

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Authors and Affiliations

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

    Mengjia Tang, Phuong Pham, Xuan Shen & Myron F. Goodman

  2. Department of Chemistry, Washington University, St. Louis, 63130, Missouri, USA

    John-Stephen Taylor

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

    Mike O'Donnell

  4. 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. Mengjia Tang
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Correspondence to Myron F. Goodman.

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Tang, M., Pham, P., Shen, X. et al. Roles of E. coli DNA polymerases IV and V in lesion-targeted and untargeted SOS mutagenesis. Nature 404, 1014–1018 (2000). https://doi.org/10.1038/35010020

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