title: Ufd1-Npl4 Recruit Cdc48 for Disassembly of Ubiquitylated CMG Helicase at the End of Chromosome Replication.
abstract: Disassembly of the Cdc45-MCM-GINS (CMG) DNA helicase is the key regulated step during DNA replication termination in eukaryotes, involving ubiquitylation of the Mcm7 helicase subunit, leading to a disassembly process that requires the Cdc48 "segregase". Here, we employ a screen to identify partners of budding yeast Cdc48 that are important for disassembly of ubiquitylated CMG helicase at the end of chromosome replication. We demonstrate that the ubiquitin-binding Ufd1-Npl4 complex recruits Cdc48 to ubiquitylated CMG. Ubiquitylation of CMG in yeast cell extracts is dependent upon lysine 29 of Mcm7, which is the only detectable site of ubiquitylation both in vitro and in vivo (though in vivo other sites can be modified when K29 is mutated). Mutation of K29 abrogates in vitro recruitment of Ufd1-Npl4-Cdc48 to the CMG helicase, supporting a model whereby Ufd1-Npl4 recruits Cdc48 to ubiquitylated CMG at the end of chromosome replication, thereby driving the disassembly reaction.
authors.affiliation: MRC Protein Phosphorylation and Ubiquitylation Unit, Sir James Black Centre, School of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, UK.
authors.forename: Marija
authors.initials: M
authors.lastname: Maric
authors.affiliation: MRC Protein Phosphorylation and Ubiquitylation Unit, Sir James Black Centre, School of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, UK.
authors.forename: Progya
authors.initials: P
authors.lastname: Mukherjee
authors.affiliation: Gene Regulation and Expression Division, School of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, UK.
authors.forename: Michael H
authors.initials: MH
authors.lastname: Tatham
authors.affiliation: Gene Regulation and Expression Division, School of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, UK.
authors.forename: Ronald
authors.initials: R
authors.lastname: Hay
authors.affiliation: MRC Protein Phosphorylation and Ubiquitylation Unit, Sir James Black Centre, School of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, UK. Electronic address: kpmlabib@dundee.ac.uk.
authors.forename: Karim
authors.initials: K
authors.lastname: Labib
mesh_terms: D015825:Chromosomes, Fungal / Q000378:metabolism*; D004261:DNA Replication; D008239:Lysine / Q000378:metabolism; D021122:Protein Subunits / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D054875:Ubiquitination
chemical_list: D021122:Protein Subunits; D029701:Saccharomyces cerevisiae Proteins; D008239:Lysine
keywords: CMG helicase; Cdc48; DNA replication termination; Ufd1-Npl4; disassembly; replisome; ubiquitin; ubiquitylation