Live-cell imaging reveals sustained centromere binding of CENP-T via CENP-A and CENP-B.
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.
Hellwig
Daniela
D
Leibniz-Institute for Age Research - Fritz Lipmann Institute, Dept. of Molecular Biology, Beutenbergstr. 11, D-07745 Jena, Germany.
Münch
Sandra
S
Orthaus
Sandra
S
Hoischen
Christian
C
Hemmerich
Peter
P
Diekmann
Stephan
S
D001324:Autoantigens / Q000378:metabolism*; D001483:Base Sequence; D002453:Cell Cycle; D002503:Centromere / Q000378:metabolism* / Q000648:ultrastructure; D000076247:Centromere Protein A; D051936:Centromere Protein B / Q000378:metabolism*; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism*; D017931:DNA Primers / Q000235:genetics; D036681:Fluorescence Recovery After Photobleaching; D031541:Fluorescence Resonance Energy Transfer; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D018386:Kinetochores / Q000378:metabolism / Q000648:ultrastructure; D008856:Microscopy, Fluorescence; D055620:Optical Phenomena; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013050:Spectrometry, Fluorescence
D001324:Autoantigens; C000619700:CENPA protein, human; C497926:CENPB protein, human; C534001:CENPT protein, human; D000076247:Centromere Protein A; D051936:Centromere Protein B; D002868:Chromosomal Proteins, Non-Histone; D017931:DNA Primers; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins