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 AbendrothStephan Diekmann

Abstract

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 nucleos...Continue Reading

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Citations

Feb 1, 2017·Nature Reviews. Genetics·Sebastian Müller, Geneviève Almouzni
Jun 11, 2016·Genes & Development·Lisa PrendergastGeneviève Almouzni

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Methods Mentioned

BETA
FRET
Fluorescence Recovery After Photobleaching
electrophoresis
Fluorescence
Fluorescence Cross
acetylation
two-hybrid
FCS
PCR
Destination

Software Mentioned

Origin
Excel

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