Replication profile from total DNA

ABSTRACT: DNA duplication is intimately connected to setting up post-replicative chromosome structures and events, but molecular details of this coordination are not well understood. A striking example occurs during yeast meiosis, where replication locally influences timing of the DNA double-strand breaks (DSBs) that initiate recombination. We show here that replication-DSB coordination is eliminated by overexpressing Dbf4-dependent Cdc7 kinase (DDK) or removing Tof1 or Csm3, components of the replication fork protection complex (FPC). DDK physically associates with Tof1, and Tof1 is dispensable for replication-DSB coordination if DDK is artificially tethered to replisomes. Furthermore, DDK phosphorylation of the DSB-promoting factor Mer2 is locally coordinated with replication, dependent on Tof1. These findings indicate that DDK recruited by FPC to replisomes phosphorylates chromatin-bound Mer2 in the wake of the replication fork, thus synchronizing replication with an early prerequisite for DSB formation. This may be a general mechanism to ensure spatial and temporal coordination of replication with other chromosomal processes. Forty-eight samples total: 8 time points from WT ARS+, WT arsM-bM-^HM-^F, DDK OP ARS+, DDK OP arsM-bM-^HM-^F,tof1M-bM-^HM-^F ARS+,tof1M-bM-^HM-^F arsM-bM-^HM-^F strains

ORGANISM(S): Saccharomyces cerevisiae

SUBMITTER: Bic MSKCC 

PROVIDER: E-GEOD-52987 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress