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HJURP is a CENP-A chromatin assembly factor sufficient to form a functional de novo kinetochore

2011; Rockefeller University Press; Volume: 194; Issue: 2 Linguagem: Inglês

10.1083/jcb.201012017

1540-8140

Meghan Barnhart, P. Henning J. L. Kuich, Madison E. Stellfox, Jared A. Ward, Emily Bassett, Ben E. Black, Daniel R. Foltz,

Genomics and Chromatin Dynamics

Centromeres of higher eukaryotes are epigenetically marked by the centromere-specific CENP-A nucleosome. New CENP-A recruitment requires the CENP-A histone chaperone HJURP. In this paper, we show that a LacI (Lac repressor) fusion of HJURP drove the stable recruitment of CENP-A to a LacO (Lac operon) array at a noncentromeric locus. Ectopically targeted CENP-A chromatin at the LacO array was sufficient to direct the assembly of a functional centromere as indicated by the recruitment of the constitutive centromere-associated network proteins, the microtubule-binding protein NDC80, and the formation of stable kinetochore–microtubule attachments. An amino-terminal fragment of HJURP was able to assemble CENP-A nucleosomes in vitro, demonstrating that HJURP is a chromatin assembly factor. Furthermore, HJURP recruitment to endogenous centromeres required the Mis18 complex. Together, these data suggest that the role of the Mis18 complex in CENP-A deposition is to recruit HJURP and that the CENP-A nucleosome assembly activity of HJURP is responsible for centromeric chromatin assembly to maintain the epigenetic mark.