The CENP-T/-W complex is a binding partner of the histone chaperone FACT.
The CENP-T/-W histone fold complex, as an integral part of the inner kinetochore, is essential for building a proper kinetochore at the centromere in order to direct chromosome segregation during mitosis. Notably, CENP-T/-W is not inherited at centromeres, and new deposition is absolutely required at each cell cycle for kinetochore function. However, the mechanisms underlying this new deposition of CENP-T/-W at centromeres are unclear. Here, we found that CENP-T deposition at centromeres is uncoupled from DNA synthesis. We identified Spt16 and SSRP1, subunits of the H2A-H2B histone chaperone facilitates chromatin transcription (FACT), as CENP-W binding partners through a proteomic screen. We found that the C-terminal region of Spt16 binds specifically to the histone fold region of CENP-T/-W. Furthermore, depletion of Spt16 impairs CENP-T and CENP-W deposition at endogenous centromeres, and site-directed targeting of Spt16 alone is sufficient to ensure local de novo CENP-T accumulation. We propose a model in which the FACT chaperone stabilizes the soluble CENP-T/-W complex in the cell and promotes dynamics of exchange, enabling CENP-T/-W deposition at centromeres.
UMR3664, Centre National de la Recherche Scientifique, Institut Curie, PSL (Paris Sciences et Lettres) Research University, F-75005 Paris, France; UMR3664, Centre National de la Recherche Scientifique, University Pierre and Marie Curie Paris 06, Sorbonne Universités, F-75005 Paris, France;
Lisa
L
Prendergast
UMR3664, Centre National de la Recherche Scientifique, Institut Curie, PSL (Paris Sciences et Lettres) Research University, F-75005 Paris, France; UMR3664, Centre National de la Recherche Scientifique, University Pierre and Marie Curie Paris 06, Sorbonne Universités, F-75005 Paris, France;
Sebastian
S
Müller
Structural Biology Program, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA;
Yiwei
Y
Liu
Structural Biology Program, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA;
Hongda
H
Huang
UMR3664, Centre National de la Recherche Scientifique, Institut Curie, PSL (Paris Sciences et Lettres) Research University, F-75005 Paris, France; Laboratoire de Spectrométrie de Masse Protéomique, Institut Curie, PSL (Paris Sciences et Lettres) Research University Centre de Recherche, Paris 75005, France;
Florent
F
Dingli
UMR3664, Centre National de la Recherche Scientifique, Institut Curie, PSL (Paris Sciences et Lettres) Research University, F-75005 Paris, France; Laboratoire de Spectrométrie de Masse Protéomique, Institut Curie, PSL (Paris Sciences et Lettres) Research University Centre de Recherche, Paris 75005, France;
Damarys
D
Loew
UMR3664, Centre National de la Recherche Scientifique, Institut Curie, PSL (Paris Sciences et Lettres) Research University, F-75005 Paris, France; UMR3664, Centre National de la Recherche Scientifique, University Pierre and Marie Curie Paris 06, Sorbonne Universités, F-75005 Paris, France;
Isabelle
I
Vassias
Structural Biology Program, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA;
Dinshaw J
DJ
Patel
Centre for Chromosome Biology, School of Natural Sciences, National University of Ireland, Galway, Ireland.
Kevin F
KF
Sullivan
UMR3664, Centre National de la Recherche Scientifique, Institut Curie, PSL (Paris Sciences et Lettres) Research University, F-75005 Paris, France; UMR3664, Centre National de la Recherche Scientifique, University Pierre and Marie Curie Paris 06, Sorbonne Universités, F-75005 Paris, France;
Geneviève
G
Almouzni
D000818:Animals; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D002503:Centromere / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006609:High Mobility Group Proteins / Q000235:genetics / Q000378:metabolism*; D056488:Histone Chaperones / Q000378:metabolism*; D006801:Humans; D018386:Kinetochores / Q000378:metabolism*; D011485:Protein Binding; D000072417:Protein Domains; D017510:Protein Folding; D040901:Proteomics; D011994:Recombinant Proteins / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D035602:Transcriptional Elongation Factors / Q000235:genetics / Q000378:metabolism*
C534001:CENPT protein, human; C534000:CENPW protein, human; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D006609:High Mobility Group Proteins; D056488:Histone Chaperones; D011994:Recombinant Proteins; C073537:SSRP1 protein, human; C489489:SUPT16H protein, human; D014157:Transcription Factors; D035602:Transcriptional Elongation Factors
CENP; centromere; histone chaperone; mitosis