The CENP-T/-W complex is a binding partner of the histone chaperone FACT

• Published in: Genes & development 2016-06, Vol.30 (11), p.1313-1326
• Main Authors: Prendergast, Lisa, Müller, Sebastian, Liu, Yiwei, Huang, Hongda, Dingli, Florent, Loew, Damarys, Vassias, Isabelle, Patel, Dinshaw J, Sullivan, Kevin F, Almouzni, Geneviève
• Format: Article
• Language: English
• Subjects: Animals; Cell Cycle Proteins - genetics; Cell Cycle Proteins - metabolism; Cell Line; Centromere - metabolism; Chromosomal Proteins, Non-Histone - metabolism; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Genetics; HeLa Cells; High Mobility Group Proteins - genetics; High Mobility Group Proteins - metabolism; Histone Chaperones - metabolism; Humans; Kinetochores - metabolism; Life Sciences; Protein Binding; Protein Domains; Protein Folding; Proteomics; Recombinant Proteins - metabolism; Research Paper; Transcription Factors - genetics; Transcription Factors - metabolism; Transcriptional Elongation Factors - genetics; Transcriptional Elongation Factors - metabolism
• ISSN: 0890-9369; 1549-5477
• DOI: 10.1101/gad.275073.115

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.