PMID:10801133
| Citation |
Tang, M, Pham, P, Shen, X, Taylor, JS, O'Donnell, M, Woodgate, R and Goodman, MF (2000) Roles of E. coli DNA polymerases IV and V in lesion-targeted and untargeted SOS mutagenesis. Nature 404:1014-8 |
|---|---|
| Abstract |
The expression of the Escherichia coli DNA polymerases pol V (UmuD'2C complex) and pol IV (DinB) increases in response to DNA damage. 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 repair. 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 beta,gamma-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 beta,gamma-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 beta,gamma-complex in the presence of non-hydrolysable ATPgammaS, indicating that an intact RecA filament may be required for translesion synthesis. |
| Links |
PubMed Online version:10.1038/35010020 |
| Keywords |
DNA Damage; DNA Polymerase beta/metabolism; DNA, Bacterial/biosynthesis; DNA, Bacterial/metabolism; DNA-Directed DNA Polymerase/metabolism; Escherichia coli/enzymology; Escherichia coli/genetics; Escherichia coli Proteins; Mutagenesis; Rec A Recombinases/metabolism; SOS Response (Genetics); Templates, Genetic |
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