The S phase checkpoint promotes the Smc5/6 complex dependent SUMOylation of Pol2, the catalytic subunit of DNA polymerase ε

English version

Autoři: Alicja Winczura ^aff001; Rowin Appanah ^aff001; Michael H. Tatham ^aff002; Ronald T. Hay ^aff002; Giacomo De Piccoli ^aff001
Působiště autorů: Warwick Medical School, University of Warwick, Coventry, United Kingdom ^aff001; Centre for Gene Regulation and Expression, School of Life Sciences, University of Dundee, United Kingdom ^aff002
Vyšlo v časopise: The S phase checkpoint promotes the Smc5/6 complex dependent SUMOylation of Pol2, the catalytic subunit of DNA polymerase ε. PLoS Genet 15(11): e32767. doi:10.1371/journal.pgen.1008427
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.pgen.1008427

Souhrn

Replication fork stalling and accumulation of single-stranded DNA trigger the S phase checkpoint, a signalling cascade that, in budding yeast, leads to the activation of the Rad53 kinase. Rad53 is essential in maintaining cell viability, but its targets of regulation are still partially unknown. Here we show that Rad53 drives the hyper-SUMOylation of Pol2, the catalytic subunit of DNA polymerase ε, principally following replication forks stalling induced by nucleotide depletion. Pol2 is the main target of SUMOylation within the replisome and its modification requires the SUMO-ligase Mms21, a subunit of the Smc5/6 complex. Moreover, the Smc5/6 complex co-purifies with Pol ε, independently of other replisome components. Finally, we map Pol2 SUMOylation to a single site within the N-terminal catalytic domain and identify a SUMO-interacting motif at the C-terminus of Pol2. These data suggest that the S phase checkpoint regulate Pol ε during replication stress through Pol2 SUMOylation and SUMO-binding ability

Klíčová slova:

DNA damage – DNA replication – Immune system proteins – Immunoblotting – Immunoprecipitation – SUMOylation – Synthesis phase – G1 phase

Zdroje

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