Evidence for lesion bypass by yeast replicative DNA polymerases during DNA damage.
The enzyme ribonucleotide reductase, responsible for the synthesis of deoxyribonucleotides (dNTP), is upregulated in response to DNA damage in all organisms. In Saccharomyces cerevisiae, dNTP concentration increases approximately 6- to 8-fold in response to DNA damage. This concentration increase is associated with improved tolerance of DNA damage, suggesting that translesion DNA synthesis is more efficient at elevated dNTP concentration. Here we show that in a yeast strain with all specialized translesion DNA polymerases deleted, 4-nitroquinoline oxide (4-NQO) treatment increases mutation frequency approximately 3-fold, and that an increase in dNTP concentration significantly improves the tolerance of this strain to 4-NQO induced damage. In vitro, under single-hit conditions, the replicative DNA polymerase epsilon does not bypass 7,8-dihydro-8-oxoguanine lesion (8-oxoG, one of the lesions produced by 4-NQO) at S-phase dNTP concentration, but does bypass the same lesion with 19-27% efficiency at DNA-damage-state dNTP concentration. The nucleotide inserted opposite 8-oxoG is dATP. We propose that during DNA damage in S. cerevisiae increased dNTP concentration allows replicative DNA polymerases to bypass certain DNA lesions.
Sabouri
Nasim
N
Department of Medical Biochemistry and Biophysics, Umeå University, SE 901 87 Umeå, Sweden.
Viberg
Jörgen
J
Goyal
Dinesh Kumar
DK
Johansson
Erik
E
Chabes
Andrei
A
D015112:4-Nitroquinoline-1-oxide / Q000633:toxicity; D004249:DNA Damage; D004257:DNA Polymerase II / Q000378:metabolism; D004261:DNA Replication; D004259:DNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism*; D003854:Deoxyribonucleotides / Q000378:metabolism; D017353:Gene Deletion; D006147:Guanine / Q000031:analogs & derivatives / Q000737:chemistry; D009154:Mutation; D015363:Quinolones / Q000633:toxicity; D012262:Ribonucleoside Diphosphate Reductase / Q000378:metabolism; D012264:Ribonucleotide Reductases / Q000378:metabolism; D016196:S Phase; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*
C120195:4-nitroquinolone-1-oxide; D003854:Deoxyribonucleotides; D015363:Quinolones; D015112:4-Nitroquinoline-1-oxide; C024829:8-hydroxyguanine; D006147:Guanine; D012264:Ribonucleotide Reductases; D012262:Ribonucleoside Diphosphate Reductase; D004257:DNA Polymerase II; D004259:DNA-Directed DNA Polymerase
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