An Anti-Checkpoint Activity for Rif1 英文参考文献.docVIP

Perspective AnAnti-CheckpointActivityforRif1 YanivHarari.,LindaRubinstein.,MartinKupiec* DepartmentofMolecularMicrobiologyandBiotechnology,TelAvivUniversity,RamatAviv,Israel 36uC) telomeres become uncapped and DNA resecting factors such as Sgs1 and Exo1arerecruited,generatingssDNA[7]. The authors followed the recruitment of thevarious factors,as wellas thebinding of checkpoint proteins, by chromatin immuno-precipitation(ChIP)attelomeric, subtelomeric, and unrelated sequences after transfer of the cells tothe restrictive temperature. cells respond to the presence of ssDNA when itslevelreaches6%–10%(at27uC) butnotatlowssDNAlevels(e.g.,at25uC), cdc13-1rif1Dcellsalreadyarrestinthecell cycle in the presence of only 2% ssDNA (at25uC). IfRif1setsthethresholdfortheDDR, then overexpression of the protein might elevatethethreshold:indeed,cdc13-1cells overexpressing Rif1 were able to grow at 29uC,aneffectsimilartotheoneobtained Chromosomal double-strand breaks (DSBs)areamongthemostseverelesions a cell has to deal with: if left unrepaired, they may lead to cell death or cancer. Thus, efficient mechanisms have evolved that respond to the presence of DSBs. These are collectively called the ‘‘DNA damage response’’ (DDR), or the ‘‘DNA damage checkpoint’’. As a result of intensivestudiesbymanyresearchgroups in several model organisms, the basic mechanismsthatrespondtoDNAdamage have been delineated: following the for- mation of DSBs, the broken ends are resected, exposing single-stranded DNA (ssDNA) which gets covered by Replica- tion Protein A (RPA), eliciting cell cycle arrest through a complex cascade of protein recruitment and phosphorylation in which several kinases take part (re- Asexpected,onceresectionbySgs1and Exo1started,theamountofRap1bound to the telomeric sequences diminished (as Rap1bindsdsDNA);however,surprising- by deleting checkpoint components such asRAD24RAD17andRAD17RAD9[8].Thus, Rif1over-expressionhasthesameeffectas a checkpoint knockout, abrogating cell