Centromeres define the chromosomal position where kinetochores form to link the chromosome to microtubules during mitosis and meiosis. Centromere identity is determined by incorporation of a specific histone H3 variant termed CenH3. As for other histones, escort and deposition of CenH3 must be ensured by histone chaperones, which handle the non-nucleosomal CenH3 pool and replenish CenH3 chromatin in dividing cells. Here, we show that the Arabidopsis orthologue of the mammalian NUCLEAR AUTOANTIGENIC SPERM PROTEIN (NASP) and Schizosaccharomyces pombe histone chaperone Sim3 is a soluble nuclear protein that binds the histone variant CenH3 and affects its abundance at the centromeres. NASPSIM3 is co-expressed with Arabidopsis CenH3 in dividing cells and binds directly to both the N-terminal tail and the histone fold domain of non-nucleosomal CenH3. Reduced NASPSIM3 expression negatively affects CenH3 deposition, identifying NASPSIM3 as a CenH3 histone chaperone.

Department of Plants and Crops Unit HortiCell Faculty of Bioscience Engineering Ghent University Coupure links 653 9000 Ghent Belgium

GReD Université Clermont Auvergne CNRS INSERM BP 38 63001 Clermont Ferrand France

Gregor Mendel Institute Vienna 1030 Austria

Institute of Molecular Biotechnology Vienna 1030 Austria

Institute of Molecular Pathology Vienna 1030 Austria

Leibniz Institute of Plant Genetics and Crop Plant Research Gatersleben Corrensstrasse 3 D 06466 Seeland Germany

Mendel Centre for Plant Genomics and Proteomics CEITEC Masaryk University Brno CZ 62500 Czech Republic

School of Biosciences University of Birmingham Edgbaston Birmingham B15 2TT UK

The Czech Academy of Sciences Institute of Experimental Botany Centre of the Region Haná for Biotechnological and Agricultural Research Šlechtitelů 31 78 371 Olomouc Czech Republic

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