Dimerization of the largest subunit of chromatin assembly factor 1: importance in vitro and during Xenopus early development.
To date, the in vivo importance of chromatin assembly factors during development in vertebrates is unknown. Chromatin assembly factor 1 (CAF-1) represents the best biochemically characterized factor promoting chromatin assembly during DNA replication or repair in human cell-free systems. Here, we identify a Xenopus homologue of the largest subunit of CAF-1 (p150). Novel dimerization properties are found conserved in both Xenopus and human p150. A region of 36 amino acids required for p150 dimerization was identified. Deletion of this domain abolishes the ability of p150 to promote chromatin assembly in vitro. A dominant-negative interference based on these dimerization properties occurs both in vitro and in vivo. In the embryo, nuclear organization was severely affected and cell cycle progression was impaired during the rapid early cleaving stages of Xenopus development. We propose that the rapid proliferation at early developmental stages necessitates the unique properties of an assembly factor that can ensure a tight coupling between DNA replication or repair and chromatin assembly.
Quivy
J P
JP
Laboratoire de Dynamique Nucléaire et Plasticité du Génome (UMR 218 du CNRS), Institut Curie/Section de Recherche, 26 rue d'Ulm, 75231 Paris Cedex 05, France.
Grandi
P
P
Almouzni
G
G
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002843:Chromatin / Q000378:metabolism*; D056504:Chromatin Assembly Factor-1; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins / Q000378:metabolism*; D019281:Dimerization; D004625:Embryo, Nonmammalian; D019143:Evolution, Molecular; D008969:Molecular Sequence Data; D009707:Nucleosomes / Q000378:metabolism; D011485:Protein Binding; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D014157:Transcription Factors; D020798:Two-Hybrid System Techniques; D014981:Xenopus / Q000196:embryology*
C533593:CHAF1B protein, human; C489780:CNOT8 protein, human; D002843:Chromatin; D056504:Chromatin Assembly Factor-1; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D009707:Nucleosomes; D014157:Transcription Factors