Human DNA polymerase ? (pol ?) can replicate across UV-induced pyrimidine dimers, and defects in the gene encoding pol ? result in a syndrome called xeroderma pigmentosum variant (XP-V). XP-V patients are prone to the development of cancer in sun-exposed areas, and cells derived from XP-V patients demonstrate increased sensitivity to UV radiation and a higher mutation rate compared with wild-type cells. pol ? has been shown to replicate across a wide spectrum of DNA lesions introduced by environmental or chemotherapeutic agents, or during nucleotide starvation, suggesting that the biological roles for pol ? are not limited to repair of UV-damaged DNA. The high error rate of pol ? requires that its intracellular activity be tightly regulated. Here, we show that the phosphorylation of pol ? increased after UV irradiation, and that treatment with caffeine, siRNA against ATR, or an inhibitor of PKC (calphostin C), reduced the accumulation of pol ? at stalled replication forks after UV irradiation or treatment with cisplatin and gemcitabine. Site-specific mutagenesis (S587A and T617A) of pol ? at two putative PKC phosphorylation sites located in the protein-protein interaction domain prevented nuclear foci formation induced by UV irradiation or treatment with gemcitabine/cisplatin. In addition, XP-V cell lines stably expressing either the S587A or T617A mutant form of pol ? were more sensitive to UV radiation and gemcitabine/cisplatin than control cells expressing wild-type pol ?. These results suggest that phosphorylation is one mechanism by which the cellular activity of pol ? is regulated. ?2008 by The National Academy of Sciences of the USA.
Date:
2008-10-28
Relation:
Proceedings of the National Academy of Sciences of the United States of America. 2008 Oct 28;105(43):16578-16583.
