title: Ctf4 Is a Hub in the Eukaryotic Replisome that Links Multiple CIP-Box Proteins to the CMG Helicase.
abstract: Replisome assembly at eukaryotic replication forks connects the DNA helicase to DNA polymerases and many other factors. The helicase binds the leading-strand polymerase directly, but is connected to the Pol α lagging-strand polymerase by the trimeric adaptor Ctf4. Here, we identify new Ctf4 partners in addition to Pol α and helicase, all of which contain a "Ctf4-interacting-peptide" or CIP-box. Crystallographic analysis classifies CIP-boxes into two related groups that target different sites on Ctf4. Mutations in the CIP-box motifs of the Dna2 nuclease or the rDNA-associated protein Tof2 do not perturb DNA synthesis genome-wide, but instead lead to a dramatic shortening of chromosome 12 that contains the large array of rDNA repeats. Our data reveal unexpected complexity of Ctf4 function, as a hub that connects multiple accessory factors to the replisome. Most strikingly, Ctf4-dependent recruitment of CIP-box proteins couples other processes to DNA synthesis, including rDNA copy-number regulation.
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: Fabrizio
authors.initials: F
authors.lastname: Villa
authors.affiliation: Department of Biochemistry, University of Cambridge, 80, Tennis Court Road, Cambridge CB2 1GA, UK.
authors.forename: Aline C
authors.initials: AC
authors.lastname: Simon
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: Maria Angeles
authors.initials: MA
authors.lastname: Ortiz Bazan
authors.affiliation: Department of Biochemistry, University of Cambridge, 80, Tennis Court Road, Cambridge CB2 1GA, UK.
authors.forename: Mairi L
authors.initials: ML
authors.lastname: Kilkenny
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, UK.
authors.forename: David
authors.initials: D
authors.lastname: Wirthensohn
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: Mel
authors.initials: M
authors.lastname: Wightman
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, UK.
authors.forename: Dijana
authors.initials: D
authors.lastname: Matak-Vinkovíc
authors.affiliation: Department of Biochemistry, University of Cambridge, 80, Tennis Court Road, Cambridge CB2 1GA, UK. Electronic address: lp212@cam.ac.uk.
authors.forename: Luca
authors.initials: L
authors.lastname: Pellegrini
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: D001665:Binding Sites; D015825:Chromosomes, Fungal / Q000201:enzymology* / Q000235:genetics; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism*; D004256:DNA Polymerase I / Q000378:metabolism; D004271:DNA, Fungal / Q000096:biosynthesis* / Q000235:genetics; D004275:DNA, Ribosomal / Q000096:biosynthesis* / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D018628:Gene Dosage; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D046912:Multiprotein Complexes; D009154:Mutation; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D016196:S Phase; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D013329:Structure-Activity Relationship
chemical_list: C078338:CTF4 protein, S cerevisiae; D004271:DNA, Fungal; D004275:DNA, Ribosomal; D004268:DNA-Binding Proteins; D047908:Intracellular Signaling Peptides and Proteins; D046912:Multiprotein Complexes; D029701:Saccharomyces cerevisiae Proteins; C119637:TOF2 protein, S cerevisiae; D004256:DNA Polymerase I; D004265:DNA Helicases; C410222:DNA2 protein, S cerevisiae