Acceptor-photobleaching FRET analysis of core kinetochore and NAC proteins in living human cells

Eur Biophys J. 2009 Jul;38(6):781-91. doi: 10.1007/s00249-009-0498-x. Epub 2009 Jun 17.

Abstract

Faithful chromatin segregation is mediated and controlled by the kinetochore protein network which assembles at centromeres. In this study, the neighbourhood relations of inner kinetochore and nucleosome-associated complex (NAC) proteins were analysed in living human interphase cells by acceptor photobleaching FRET. The data indicate that CENP-U is in close vicinity to CENP-I as well as to CENP-B and that CENP-M is close to CENP-T.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics
  • Blotting, Western
  • Cell Line, Tumor
  • Centromere Protein B / chemistry
  • Centromere Protein B / metabolism
  • Chromosomal Proteins, Non-Histone / chemistry
  • Chromosomal Proteins, Non-Histone / metabolism
  • Fluorescence Resonance Energy Transfer / methods
  • Green Fluorescent Proteins / genetics
  • Humans
  • Interphase / physiology
  • Kinetochores / chemistry*
  • Kinetochores / metabolism
  • Luminescent Proteins / genetics
  • Nuclear Proteins / chemistry*
  • Nuclear Proteins / metabolism
  • Photobleaching
  • Transfection

Substances

  • Bacterial Proteins
  • CENPB protein, human
  • CENPT protein, human
  • Centromere Protein B
  • Chromosomal Proteins, Non-Histone
  • Luminescent Proteins
  • Nuclear Proteins
  • centromere protein C
  • enhanced green fluorescent protein
  • yellow fluorescent protein, Bacteria
  • Green Fluorescent Proteins