Live-cell imaging reveals sustained centromere binding of CENP-T via CENP-A and CENP-B

• Published in: Journal of biophotonics Vol. 1; no. 3; pp. 245 - 254
• Main Authors: Hellwig, D., Münch, S., Orthaus, S., Hoischen, C., Hemmerich, P., Diekmann, S.
• Format: Journal Article
• Language: English
• Published: Berlin WILEY-VCH Verlag 01.08.2008; WILEY‐VCH Verlag
• Subjects: Autoantigens - metabolism; Base Sequence; Cell Cycle; centromere; Centromere - metabolism; Centromere - ultrastructure; Centromere Protein A; Centromere Protein B - metabolism; Chromosomal Proteins, Non-Histone - genetics; Chromosomal Proteins, Non-Histone - metabolism; DNA Primers - genetics; FCS; Fluorescence Recovery After Photobleaching; Fluorescence Resonance Energy Transfer; FRAP; FRET; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; HeLa Cells; Humans; kinetochore; Kinetochores - metabolism; Kinetochores - ultrastructure; live-cell imaging; Microscopy, Fluorescence; mitosis; NAC complex; Optical Phenomena; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Spectrometry, Fluorescence
• ISSN: 1864-063X; 1864-0648
• DOI: 10.1002/jbio.200810014

At the centromere, a network of proteins, the kinetochore, assembles in order to grant correct chromatin segregation. In this study the dynamics and molecular interactions of the inner kinetochore protein CENP‐T were analyzed employing a variety of fluorescence microscopy techniques in living human cells. Acceptor‐bleaching FRET indicates that CENP‐T directly associates with CENP‐A and CENP‐B. CENP‐T exchange into centromeres is restricted to the S‐phase of the cell cycle as revealed by FRAP, suggesting a coreplicational loading mechanism, as we have recently also demonstrated for CENP‐I. These properties make CENP‐T one of the basic inner kinetochore proteins with most further proteins binding downstream, suggesting a fundamental role of CENP‐T in kinetochore function. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)