Replication past O(6)-methylguanine by yeast and human DNA polymerase eta.
O(6)-Methylguanine (m6G) is formed by the action of alkylating agents such as N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) on DNA. m6G is a highly mutagenic and carcinogenic lesion, and it presents a block to synthesis by DNA polymerases. Here, we provide genetic and biochemical evidence for the involvement of yeast and human DNA polymerase eta (Poleta) in the replicative bypass of m6G lesions in DNA. The formation of MNNG-induced mutations is almost abolished in the rad30Delta pol32Delta double mutant of yeast, which lacks the RAD30 gene that encodes Poleta and the Pol32 subunit of DNA polymerase delta (Poldelta). Although Poldelta can function in the mutagenic bypass of m6G lesions, our biochemical studies indicate that Poleta is much more efficient in replicating through m6G than Poldelta. Both Poleta and Poldelta insert a C or a T residue opposite from m6G; Poleta, however, is more accurate, as it inserts a C about twice as frequently as Poldelta. Alkylating agents are used in the treatment of malignant tumors, including lymphomas, brain tumors, melanomas, and gastrointestinal carcinomas, and the clinical effectiveness of these agents derives at least in part from their ability to form m6G in DNA. Inactivation of Poleta could afford a useful strategy for enhancing the effectiveness of these agents in cancer chemotherapy.
Haracska
L
L
University of Texas Medical Branch, Sealy Center for Molecular Science, Galveston, Texas 77555-1061, USA.
Prakash
S
S
Prakash
L
L
D000477:Alkylating Agents / Q000494:pharmacology; D001483:Base Sequence; D003596:Cytosine / Q000378:metabolism; D004258:DNA Polymerase III / Q000235:genetics / Q000378:metabolism; D004260:DNA Repair; D004261:DNA Replication; D004259:DNA-Directed DNA Polymerase / Q000235:genetics* / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005656:Fungal Proteins / Q000378:metabolism; D006147:Guanine / Q000031:analogs & derivatives* / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008769:Methylnitronitrosoguanidine / Q000494:pharmacology; D008969:Molecular Sequence Data; D009154:Mutation; D013936:Thymidine / Q000378:metabolism; D000094769:DNA Polymerase iota; D000099270:Y-Family DNA Polymerases
D000477:Alkylating Agents; D003596:Cytosine; D004258:DNA Polymerase III; D004259:DNA-Directed DNA Polymerase; D005656:Fungal Proteins; D006147:Guanine; D008769:Methylnitronitrosoguanidine; D013936:Thymidine; D000094769:DNA Polymerase iota; C008449:O-(6)-methylguanine; C112338:Rad30 protein; D000099270:Y-Family DNA Polymerases; C000722070:POLI protein, human