Centromere targeting of Mis18 requires the interaction with DNA and H2A-H2B in fission yeast

Cell Mol Life Sci. 2021 Jan;78(1):373-384. doi: 10.1007/s00018-020-03502-1. Epub 2020 Apr 21.

Abstract

Faithful chromosome segregation during mitosis requires the correct assembly of kinetochore on the centromere. CENP-A is a variant of histone H3, which specializes the centromere region on chromatin and mediates the kinetochore assembly. The Mis18 complex plays a critical role in initiating the centromere loading of the newly-synthesized CENP-A. However, it remains unclear how Mis18 complex (spMis18, spMis16 and spMis19) is located to the centromere to license the recruitment of Cnp1CENP-A in Schizosaccharomyces pombe. We found that spMis18 directly binds to nucleosomal DNA through its extreme C-terminus and interacts with H2A-H2B dimer via the acidic region on the surface of its Yippee-like domain. Live-cell imaging confirmed that mutation of the acidic region and deletion of the extreme C-terminus significantly impairs the localization of spMis18 and Cnp1 to the centromere and delays chromosome segregation during mitosis. Our findings illustrate that the interaction of spMis18 with histone H2A-H2B and DNA plays important roles in the recruitment of spMis18 and Cnp1 to the centromere in fission yeast.

Keywords: Centromere; Cnp1; DNA; Histone H2A–H2B; Mis18.

MeSH terms

  • Carrier Proteins / chemistry
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism*
  • Centromere / metabolism
  • Chromosomal Proteins, Non-Histone / genetics
  • Chromosomal Proteins, Non-Histone / metabolism
  • Chromosome Segregation
  • Crystallography, X-Ray
  • DNA / chemistry
  • DNA / metabolism*
  • Dimerization
  • Histones / genetics
  • Histones / metabolism*
  • Microscopy, Fluorescence
  • Mitosis
  • Molecular Dynamics Simulation
  • Multiprotein Complexes / chemistry
  • Multiprotein Complexes / genetics
  • Multiprotein Complexes / metabolism
  • Mutagenesis
  • Protein Binding
  • Protein Domains
  • Protein Structure, Tertiary
  • Schizosaccharomyces / genetics
  • Schizosaccharomyces / metabolism*
  • Schizosaccharomyces pombe Proteins / chemistry
  • Schizosaccharomyces pombe Proteins / genetics
  • Schizosaccharomyces pombe Proteins / metabolism*
  • Time-Lapse Imaging

Substances

  • Carrier Proteins
  • Chromosomal Proteins, Non-Histone
  • Cnp1 protein, S pombe
  • Histones
  • Mis18 protein, S pombe
  • Mis19 protein, S pombe
  • Multiprotein Complexes
  • Schizosaccharomyces pombe Proteins
  • DNA