dna-pk-dependent rpa2 hyperphosphorylation facilitates dna repair and suppresses sister chromatid exchangedna-pk-dependent rpa2 hyperphosphorylation促进dna修复和抑制姐妹染色单体交换.pdfVIP

DNA-PK-Dependent RPA2 Hyperphosphorylation Facilitates DNA Repair and Suppresses Sister Chromatid Exchange ¤ Hungjiun Liaw , Deokjae Lee, Kyungjae Myung* Genome Instability Section, Genetics and Molecular Biology Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland, United States of America Abstract Hyperphosphorylation of RPA2 at serine 4 and serine 8 (S4, S8) has been used as a marker for activation of the DNA damage response. What types of DNA lesions cause RPA2 hyperphosphorylation, which kinase(s) are responsible for them, and what is the biological outcome of these phosphorylations, however, have not been fully investigated. In this study we demonstrate that RPA2 hyperphosphorylation occurs primarily in response to genotoxic stresses that cause high levels of DNA double-strand breaks (DSBs) and that the DNA-dependent protein kinase complex (DNA-PK) is responsible for the modifications in vivo. Alteration of S4, S8 of RPA2 to alanines, which prevent phosphorylations at these sites, caused increased mitotic entry with concomitant increases in RAD51 foci and homologous recombination. Taken together, our results demonstrate that RPA2 hyperphosphorylation by DNA-PK in response to DSBs blocks unscheduled homologous recombination and delays mitotic entry. This pathway thus permits cells to repair DNA damage properly and increase cell viability. Citation: Liaw H, Lee D, Myung K (2011) DNA-PK-Dependent RPA2 Hyperphosphorylation Facilitates DNA Repair and Suppresses Sister Chromatid Exchange. PLoS ONE 6(6): e21424. doi:10.1371/journal.pone.0021424 Editor: David T. Kirkpatrick, University of Minnesota, United States of America Received February 2, 2011; Accepted May 30, 2011; Published June 22, 2011 This is an open-ac