The CENP-T C-Terminus Is Exclusively Proximal to H3.1 and not to H3.2 or H3.3

International Journal of Molecular Sciences

Christian Abendroth²  Antje Hofmeister²  Sandra B. Hake⁴  Paul K. Kamweru²  Elke Miess²  Carsten Dornblut²  Isabell Kﳿner²  Wen Deng¹  Heinrich Leonhardt¹  Sandra Orthaus³  Christian Hoischen²  Stephan Diekmann² 

[1] Department of Biology II, Center for Integrated Protein Science, Ludwig-Maximilians-Universität Munich, Planegg-Martinsried, D-82152 Munich, Germany; E-Mails:;Molecular Biology, Fritz Lipmann Institute, Beutenbergstr. 11, D-07745 Jena, Germany; E-Mails:;PicoQuant GmbH, Kekulestr. 7, D-12489 Berlin, Germany; E-Mail:;Department of Molecular Biology, Center for Integrated Protein Science Munich (CIPSM), Adolf-Butenandt-Institute, Ludwig-Maximilians-Universität Munich, Schillerstr. 44, D-80336 Munich, Germany; E-Mail:

关键词: centromere;    kinetochore;    Constitutive Centromere-Associated Network (CCAN);    mitosis;    histone variants;    chromatin;    Förster Resonance Energy Transfer (FRET);   

DOI  :  10.3390/ijms16035839

来源: mdpi

【 摘 要 】

The kinetochore proteins assemble onto centromeric chromatin and regulate DNA segregation during cell division. The inner kinetochore proteins bind centromeres while most outer kinetochore proteins assemble at centromeres during mitosis, connecting the complex to microtubules. The centromere–kinetochore complex contains specific nucleosomes and nucleosomal particles. CENP-A replaces canonical H3 in centromeric nucleosomes, defining centromeric chromatin. Next to CENP-A, the CCAN multi-protein complex settles which contains CENP-T/W/S/X. These four proteins are described to form a nucleosomal particle at centromeres. We had found the CENP-T C-terminus and the CENP-S termini next to histone H3.1 but not to CENP-A, suggesting that the Constitutive Centromere-Associated Network (CCAN) bridges a CENP-A- and a H3-containing nucleosome. Here, we show by in vivo FRET that this proximity between CENP-T and H3 is specific for H3.1 but neither for the H3.1 mutants H3.1^C96A and H3.1^C110A nor for H3.2 or H3.3. We also found CENP-M next to H3.1 but not to these H3.1 mutants. Consistently, we detected CENP-M next to CENP-S. These data elucidate the local molecular neighborhood of CCAN proteins next to a H3.1-containing centromeric nucleosome. They also indicate an exclusive position of H3.1 clearly distinct from H3.2, thus documenting a local, and potentially also functional, difference between H3.1 and H3.2.

【 授权许可】

CC BY   
© 2015 by the authors; licensee MDPI, Basel, Switzerland.