Roles of E. coli DNA polymerases IV and V in lesion-targeted and untargeted SOS mutagenesis.
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.
Tang
M
M
Department of Biological Sciences and Chemistry, University of Southern California, University Park, Los Angeles 90089-1340, USA.
Pham
P
P
Shen
X
X
Taylor
J S
JS
O'Donnell
M
M
Woodgate
R
R
Goodman
M F
MF
D004249:DNA Damage; D019951:DNA Polymerase beta / Q000378:metabolism*; D004269:DNA, Bacterial / Q000096:biosynthesis / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D029968:Escherichia coli Proteins; D016296:Mutagenesis; D011938:Rec A Recombinases / Q000378:metabolism; D013014:SOS Response, Genetics; D013698:Templates, Genetic
D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D011938:Rec A Recombinases; D019951:DNA Polymerase beta; C120926:DNA polymerase V, E coli; D004259:DNA-Directed DNA Polymerase