HJURP binds CENP-A via a highly conserved N-terminal domain and mediates its deposition at centromeres

Proc Natl Acad Sci U S A. 2010 Jan 26;107(4):1349-54. doi: 10.1073/pnas.0913709107. Epub 2010 Jan 6.

Abstract

The human histone H3 variant, CENP-A, replaces the conventional histone H3 in centromeric chromatin and, together with centromere-specific DNA-binding factors, directs the assembly of the kinetochore. We purified the prenucelosomal e-CENP-A complex. We found that HJURP, a member of the complex, was required for cell cycle specific targeting of CENP-A to centromeres. HJURP facilitated efficient deposition of CENP-A/H4 tetramers to naked DNA in vitro. Bacterially expressed HJURP binds at a stoichiometric ratio to the CENP-A/H4 tetramer but not to the H3/H4 tetramer. The binding occurred through a conserved HJURP short N-terminal domain, termed CBD. The novel characteristic identified in vertebrates that we named TLTY box of CBD, was essential for formation of the HJURP-CENP-A/H4 complex. Our data identified HJURP as a vertebrate CENP-A chaperone and dissected its mode of interactions with CENP-A.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Autoantigens / chemistry*
  • Autoantigens / genetics
  • Autoantigens / metabolism*
  • Cell Cycle
  • Centromere / metabolism*
  • Centromere Protein A
  • Chromosomal Proteins, Non-Histone / chemistry*
  • Chromosomal Proteins, Non-Histone / genetics
  • Chromosomal Proteins, Non-Histone / metabolism*
  • Conserved Sequence
  • DNA-Binding Proteins / chemistry*
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • HeLa Cells
  • Histones / metabolism
  • Humans
  • Models, Molecular
  • Molecular Sequence Data
  • Nucleosomes / metabolism
  • Protein Binding
  • Protein Interaction Domains and Motifs*
  • Protein Multimerization
  • Protein Structure, Quaternary
  • RNA, Small Interfering / genetics
  • Sequence Alignment

Substances

  • Autoantigens
  • CENPA protein, human
  • Centromere Protein A
  • Chromosomal Proteins, Non-Histone
  • DNA-Binding Proteins
  • HJURP protein, human
  • Histones
  • Nucleosomes
  • RNA, Small Interfering