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Concomitantly with DNA replication, the cohesin complex establishes cohesion between the newly synthesized sister chromatids. Cohesion establishment requires acetylation of the cohesin subunit Smc3 by the Eco1 acetyltransferase. Acetylation is tightly coupled to DNA replication and depends on (i) DNA replication factors, (ii) DNA structures that transiently form at replication forks, as well as (iii) auxiliary cohesin complex components. Here, we describe a protocol to study DNA replication-coupled cohesin acetylation using purified budding yeast proteins.

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Biochemical Reconstitution of Replication-Coupled Cohesin Acetylation

  • 实验方案

  • 发表日期：

    2026年

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  • Masashi Minamino,
  • Céline Bouchoux,
  • Frank Uhlmann

摘要

Concomitantly with DNA replication, the cohesin complex establishes cohesion between the newly synthesized sister chromatids. Cohesion establishment requires acetylation of the cohesin subunit Smc3 by the Eco1 acetyltransferase. Acetylation is tightly coupled to DNA replication and depends on (i) DNA replication factors, (ii) DNA structures that transiently form at replication forks, as well as (iii) auxiliary cohesin complex components. Here, we describe a protocol to study DNA replication-coupled cohesin acetylation using purified budding yeast proteins.