Responses to the major acrolein-derived deoxyguanosine adduct in Escherichia coli.
Acrolein, a reactive alpha,beta-unsaturated aldehyde found ubiquitously in the environment and formed endogenously in mammalian cells, reacts with DNA to form an exocyclic DNA adduct, 3H-8-hydroxy-3-(beta-D-2'-deoxyribofuranosyl)-5,6,7,8-tetrahydropyrido[3,2-a]purine-9-one (gamma-OH-PdG). The cellular processing and mutagenic potential of gamma-OH-PdG have been examined, using a site-specific approach in which a single adduct is embedded in double-strand plasmid DNA. Analysis of progeny plasmid reveals that this adduct is excised by nucleotide excision repair. The apparent level of inhibition of DNA synthesis is approximately 70% in Escherichia coli DeltarecA, uvrA. The block to DNA synthesis can be overcome partially by recA-dependent recombination repair. Targeted G --> T transversions were observed at a frequency of 7 x 10(-4)/translesion synthesis. Inactivation of polB, dinB, and umuD,C genes coding for "SOS" DNA polymerases did not affect significantly the efficiency or fidelity of translesion synthesis. In vitro primer extension experiments revealed that the Klenow fragment of polymerase I catalyzes error-prone synthesis, preferentially incorporating dAMP and dGMP opposite gamma-OH-PdG. We conclude from this study that DNA polymerase III catalyzes translesion synthesis across gamma-OH-PdG in an error-free manner. Nucleotide excision repair, recombination repair, and highly accurate translesion synthesis combine to protect E. coli from the potential genotoxicity of this DNA adduct.
Yang
I Y
IY
Laboratory of Chemical Biology, Department of Pharmacological Sciences, State University of New York at Stony Brook, Stony Brook, New York 11794-8651, USA.
Hossain
M
M
Miller
H
H
Khullar
S
S
Johnson
F
F
Grollman
A
A
Moriya
M
M
D000171:Acrolein / Q000378:metabolism / Q000494:pharmacology*; D001483:Base Sequence; D002384:Catalysis; D004256:DNA Polymerase I / Q000378:metabolism; D004258:DNA Polymerase III / Q000378:metabolism; D017931:DNA Primers; D004260:DNA Repair; D004261:DNA Replication / Q000187:drug effects; D004269:DNA, Bacterial / Q000096:biosynthesis / Q000187:drug effects / Q000378:metabolism; D003849:Deoxyguanosine / Q000378:metabolism*; D004926:Escherichia coli / Q000187:drug effects* / Q000235:genetics; D008969:Molecular Sequence Data; D011995:Recombination, Genetic
D017931:DNA Primers; D004269:DNA, Bacterial; D000171:Acrolein; D004256:DNA Polymerase I; D004258:DNA Polymerase III; D003849:Deoxyguanosine