A separable domain of the p150 subunit of human chromatin assembly factor-1 promotes protein and chromosome associations with nucleoli

Mol Biol Cell. 2014 Sep 15;25(18):2866-81. doi: 10.1091/mbc.E14-05-1029. Epub 2014 Jul 23.

Abstract

Chromatin assembly factor-1 (CAF-1) is a three-subunit protein complex conserved throughout eukaryotes that deposits histones during DNA synthesis. Here we present a novel role for the human p150 subunit in regulating nucleolar macromolecular interactions. Acute depletion of p150 causes redistribution of multiple nucleolar proteins and reduces nucleolar association with several repetitive element-containing loci. Of note, a point mutation in a SUMO-interacting motif (SIM) within p150 abolishes nucleolar associations, whereas PCNA or HP1 interaction sites within p150 are not required for these interactions. In addition, acute depletion of SUMO-2 or the SUMO E2 ligase Ubc9 reduces α-satellite DNA association with nucleoli. The nucleolar functions of p150 are separable from its interactions with the other subunits of the CAF-1 complex because an N-terminal fragment of p150 (p150N) that cannot interact with other CAF-1 subunits is sufficient for maintaining nucleolar chromosome and protein associations. Therefore these data define novel functions for a separable domain of the p150 protein, regulating protein and DNA interactions at the nucleolus.

MeSH terms

  • Cell Nucleolus / metabolism*
  • Chromatin Assembly Factor-1 / physiology*
  • Chromosomes, Human / metabolism
  • HeLa Cells
  • Humans
  • Ki-67 Antigen / metabolism
  • Protein Binding
  • Protein Interaction Domains and Motifs
  • Protein Transport
  • Transcription Factors

Substances

  • CNOT8 protein, human
  • Chromatin Assembly Factor-1
  • Ki-67 Antigen
  • Transcription Factors