reversal of pcna ubiquitylation by ubp10 in saccharomyces cerevisiae逆转的pcna ubiquitylation ubp10酿酒酵母.pdfVIP

Reversal of PCNA Ubiquitylation by Ubp10 in Saccharomyces cerevisiae ´ 1 ´ 1 2 2 1,3 Alfonso Gallego-Sanchez , Sonia Andres , Francisco Conde , Pedro A. San-Segundo , Avelino Bueno * ´ ´ ´ ´ 1 Instituto de Biologıa Molecular y Celular del Cancer, Universidad de Salamanca/CSIC, Salamanca, Spain, 2 Instituto de Biologıa Funcional y Genomica, Universidad de ´ ´ Salamanca/CSIC, Salamanca, Spain, 3 Departamento de Microbiologıa y Genetica, Universidad de Salamanca/CSIC, Salamanca, Spain Abstract Regulation of PCNA ubiquitylation plays a key role in the tolerance to DNA damage in eukaryotes. Although the evolutionary conserved mechanism of PCNA ubiquitylation is well understood, the deubiquitylation of ubPCNA remains poorly characterized. Here, we show that the histone H2BK123 ubiquitin protease Ubp10 also deubiquitylates ubPCNA in Saccharomyces cerevisiae. Our results sustain that Ubp10-dependent deubiquitylation of the sliding clamp PCNA normally takes place during S phase, likely in response to the simple presence of ubPCNA. In agreement with this, we show that Ubp10 forms a complex with PCNA in vivo. Interestingly, we also show that deletion of UBP10 alters in different ways the interaction of PCNA with DNA polymerase f–associated protein Rev1 and with accessory subunit Rev7. While deletion of UBP10 enhances PCNA–Rev1 interaction, it decreases significantly Rev7 binding to