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Efficiency and accuracy of SOS-induced DNA polymerases replicating benzo a pyrene-7,8-diol 9,10-epoxide A and G adducts

  1. Author:
    Shen, X.
    Sayer, J. M.
    Kroth, H.
    Ponten, I.
    O'Donnell, M.
    Woodgate, R.
    Jerina, D. M.
    Goodman, M. F.

  2. Author Address

    Univ So Calif, Dept Biol Sci, SHS Rm 172, Univ Pk, Los Angeles, CA 90089 USA. Univ So Calif, Hedco Mol Biol Lab, Dept Biol Sci & Chem, Los Angeles, CA 90089 USA. NIDDK, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. NICHHD, Sect DNA Replicat Repair & Mutagenesis, NIH, Bethesda, MD 20892 USA. NCI, Frederick Canc Res & Dev Ctr, Chem Carcinogenesis Lab, Frederick, MD 21702 USA. Rockefeller Univ, New York, NY 10021 USA. Howard Hughes Med Inst, New York, NY 10021 USA. Goodman MF Univ So Calif, Dept Biol Sci, SHS Rm 172, Univ Pk, Los Angeles, CA 90089 USA.
    1. Year: 2002
  2. Journal: Journal of Biological Chemistry
    1. 277
    2. 7
    3. Pages: 5265-5274
  4. Type of Article: Article
  1. Abstract:

    Nucleotide incorporation fidelity, mismatch extension, and translesion DNA synthesis efficiencies were determined using SOS-induced Escherichia coli DNA polymerases (pol) II, IV, and V to copy 10R and 10S isomers of trans-opened benzo[a]pyrene- 7,8-diol 9,10-epoxide (BaP DE) A and G adducts. A-BaP DE adducts were bypassed by pol V with moderate accuracy and considerably higher efficiency than by pol II or IV. Error- prone pol V copied G-BaP DE-adducted DNA poorly, forming A(.)G- BaP DE-S and -R mismatches over C(.)G-BaP DE-S and -R correct matches by factors of similar to350- and 130-fold, respectively, even favoring G(.)G-BaP DE mismatches over correct matches by factors of 2-4-fold. In contrast, pol IV bypassed G-BaP DE adducts with the highest efficiency and fidelity, making misincorporations with a frequency of 10(-2) to 10(-4) depending on sequence context. G-BaP DE-S-adducted M13 DNA yielded 4-fold fewer plaques when transfected into SOS- induced DeltadinB (pol IV-deficient) mutant cells compared with the isogenic wild-type E. coli strain, consistent with the in vitro data showing that pol IV was most effective by far at copying the G-BaP DE-S adduct. SOS polymerases are adept at copying a variety of lesions, but the relative contribution of each SOS polymerase to copying damaged DNA appears to be determined by the lesion's identity.

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