Single-molecule analysis reveals clustering and epigenetic regulation of replication origins at the yeast rDNA locus

Author:

Pasero Philippe,Bensimon Aaron,Schwob Etienne

Abstract

How eukaryotes specify their replication origins is an important unanswered question. Here, we analyze the replicative organization of yeast rDNA, which consists of ∼150 identical repeats, each containing a potential origin. Using DNA combing and single-molecule imaging, we show that functional rDNA origins are clustered and interspersed with large domains where initiation is silenced. This repression is largely mediated by the Sir2p histone-deacetylase. Increased origin firing insir2Δ mutants leads to the accumulation of circular rDNA species, a major determinant of yeast aging. We conclude that rDNA replication is regulated epigenetically and that Sir2p may promote genome stability and longevity by suppressing replication-dependent rDNA recombination.

Publisher

Cold Spring Harbor Laboratory

Subject

Developmental Biology,Genetics

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