Published online 29 July 2004
Nucleic Acids Research, Vol. 32 No. 13 © Oxford University Press 2004; all rights reserved
Role of DNA polymerase 
in the bypass of abasic sites in yeast cells
Graduate Center for Toxicology, University of Kentucky, Lexington, KY 40536, USA
* To whom correspondence should be addressed. Tel: +1 859 323 5784; Fax: +1 859 323 1059; Email: zwang{at}uky.edu
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors
Received February 7, 2004; Revised May 21, 2004; Accepted June 30, 2004
Abasic (AP) sites are major DNA lesions and are highly mutagenic. AP site-induced mutagenesis largely depends on translesion synthesis. We have examined the role of DNA polymerase 
(Pol) in translesion synthesis of AP sites by replicating a plasmid containing a site-specific AP site in yeast cells. In wild-type cells, AP site bypass resulted in preferred C insertion (62%) over A insertion (21%), as well as –1 deletion (3%), and complex event (14%) containing multiple mutations. In cells lacking Pol (rad30), Rev1, Pol
(rev3), and both Pol and Pol, translesion synthesis was reduced to 30%, 30%, 15% and 3% of the wild-type level, respectively. C insertion opposite the AP site was reduced in rad30 mutant cells and was abolished in cells lacking Rev1 or Pol, but significant A insertion was still detected in these mutant cells. While purified yeast Pol
effectively inserted an A opposite the AP site in vitro, purified yeast Pol
was much less effective in A insertion opposite the lesion due to its 3'
5' proofreading exonuclease activity. Purified yeast Pol performed extension synthesis from the primer 3' A opposite the lesion. These results show that Pol is involved in translesion synthesis of AP sites in yeast cells, and suggest that an important role of Pol is to catalyze extension following A insertion opposite the lesion. Consistent with these conclusions, rad30 mutant cells were sensitive to methyl methanesulfonate (MMS), and rev1 rad30 or rev3 rad30 double mutant cells were synergistically more sensitive to MMS than the respective single mutant strains.
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