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Updated: Jun 23, 2026

Immunofluorescence Analysis of Endogenous and Exogenous Centromere-kinetochore Proteins
Published on: March 3, 2016
Live-cell imaging reveals sustained centromere binding of CENP-T via CENP-A and CENP-B
Daniela Hellwig1, Sandra Münch, Sandra Orthaus
1Leibniz-Institute for Age Research - Fritz Lipmann Institute, Dept. of Molecular Biology, Beutenbergstr. 11, D-07745 Jena, Germany.
Abstract:
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.
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