ABSTRACT
Mono-ubiquitination of histone H2B (H2B-Ub1) is a conserved modification that plays central role in regulating numerous biological processes including the DNA damage response, gene transcription, and DNA replication. Previous studies have revealed that H2B-Ub1 promotes recovery from replication stress by mediating Rad53 phosphorylation (Rad53-P), and activation of the intra-S replication checkpoint, in order to limit fork progression, and associated DNA damage. Since such mono-ubiquitination is a reversible process, we examined the role of H2B-Ub1 deubiquitination during replication stress. Using an experimental system in yeast which mimics H2B-Ub1 accumulation, we show that cells become sensitive to the replication stress induced by HU. This stress response was accompanied by Rad53-P accumulation, and delayed recovery from intra-S checkpoint arrest. Furthermore, we show that similar effects were recapitulated by the accumulation of endogenous H2B-Ub1, induced by the co-inactivation of the deubiquitinating enzyme, Ubp10, and Spt16, a FACT histone chaperone family member. While it has been well established that H2B mono-ubiquitination plays an essential role in recovering from replication stress, our data reveal that H2B-Ub1 deubiquitination is also essential for this process.