The replication kinase Cdc7-Dbf4 promotes the interaction of the p150 subunit of chromatin assembly factor 1 with proliferating cell nuclear antigen.
The coordination of chromatin assembly with DNA replication, which is essential for genomic stability, requires the combined activation of histone deposition with the firing of replication origins. We report here the direct interaction of chromatin assembly factor 1 (CAF1), a key factor involved in histone deposition, with the replication kinase Cdc7-Dbf4. We isolated a complex containing both the largest subunit of CAF1 (p150) and the Cdc7-Dbf4 kinase specifically in S phase and thus prove the existence of this interaction in vivo. We then show that the Cdc7-Dbf4 kinase efficiently phosphorylates p150. This event induces a change in p150 oligomerization state, which promotes binding to proliferating cell nuclear antigen (PCNA). Conversely, CAF1 recruitment is reduced in a PCNA/DNA loading assay using Cdc7-depleted extracts. Our data define p150 as a new target for this kinase with implications for the coordination between DNA replication and CAF1 functions.
Gérard
Annabelle
A
Institut Curie, Section de Recherche, UMR218 du CNRS, 26 rue d'Ulm, 75248 Paris Cedex 05, France.
Koundrioukoff
Stéphane
S
Ramillon
Vincent
V
Sergère
Jean-Christophe
JC
Mailand
Niels
N
Quivy
Jean-Pierre
JP
Almouzni
Geneviève
G
D002453:Cell Cycle; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D056504:Chromatin Assembly Factor-1; D002868:Chromosomal Proteins, Non-Histone / Q000032:analysis / Q000378:metabolism*; D004261:DNA Replication / Q000235:genetics; D004268:DNA-Binding Proteins / Q000032:analysis / Q000378:metabolism*; D019281:Dimerization; D005434:Flow Cytometry; D006801:Humans; D047468:Immunoprecipitation; D051738:Origin Recognition Complex / Q000235:genetics; D010766:Phosphorylation; D018809:Proliferating Cell Nuclear Antigen / Q000032:analysis / Q000378:metabolism*; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018741:Replication Origin / Q000235:genetics; D014157:Transcription Factors
C489780:CNOT8 protein, human; D018797:Cell Cycle Proteins; D056504:Chromatin Assembly Factor-1; D002868:Chromosomal Proteins, Non-Histone; C485678:DBF4 protein, human; D004268:DNA-Binding Proteins; D051738:Origin Recognition Complex; D018809:Proliferating Cell Nuclear Antigen; D014157:Transcription Factors; C068619:CDC7 protein, human; D017346:Protein Serine-Threonine Kinases