PCNA monoubiquitylation and DNA polymerase   ubiquitin-binding domain are required to prevent 8-oxoguanine-induced mutagenesis in Saccharomyces cerevisiae

• Published in: Nucleic acids research 2009-05, Vol.37 (8), p.2549-2559
• Main Authors: van der Kemp, P. A., de Padula, M., Burguiere-Slezak, G., Ulrich, H. D., Boiteux, S.
• Format: Article
• Language: English
• Subjects: Saccharomyces cerevisiae
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/gkp105

7,8-Dihydro-8-oxoguanine (8-oxoG) is an abundant and mutagenic DNA lesion. In Saccharomyces cerevisiae, the 8-oxoG DNA N-glycosylase (Ogg1) acts as the primary defense against 8-oxoG. Here, we present evidence for cooperation between Rad18-Rad6-dependent monoubiquitylation of PCNA at K164, the damage-tolerant DNA polymerase η and the mismatch repair system (MMR) to prevent 8-oxoG-induced mutagenesis. Preventing PCNA modification at lysine 164 (pol30-K164R) results in a dramatic increase in GC to TA mutations due to endogenous 8-oxoG in Ogg1-deficient cells. In contrast, deletion of RAD5 or SIZ1 has little effect implying that the modification of PCNA relevant for preventing 8-oxoG-induced mutagenesis is monoubiquitin as opposed to polyubiquitin or SUMO. We also report that the ubiquitin-binding domain (UBZ) of Pol η is essential to prevent 8-oxoG-induced mutagenesis but only in conjunction with a functional PCNA-binding domain (PIP). We propose that PCNA is ubiquitylated during the repair synthesis reaction after the MMR-dependent excision of adenine incorporated opposite to 8-oxoG. Monoubiquitylation of PCNA would favor the recruitment of Pol η thereby allowing error-free incorporation of dCMP opposite to 8-oxoG. This study suggests that Pol η and the post-replication repair (PRR) machinery can also prevent mutagenesis at DNA lesions that do not stall replication forks.