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Abstract
The histone H3 variant CenH3, called CENP-A in humans, is central in centromeric chromatin
to ensure proper chromosome segregation. In the absence of an underlying DNA sequence,
it is still unclear how CENP-A deposition at centromeres is determined. Here, we purified
non-nucleosomal CENP-A complexes to identify direct CENP-A partners involved in such
a mechanism and identified HJURP. HJURP was not detected in H3.1- or H3.3-containing
complexes, indicating its specificity for CENP-A. HJURP centromeric localization is
cell cycle regulated, and its transient appearance at the centromere coincides precisely
with the proposed time window for new CENP-A deposition. Furthermore, HJURP downregulation
leads to a major reduction in CENP-A at centromeres and impairs deposition of newly
synthesized CENP-A, causing mitotic defects. We conclude that HJURP is a key factor
for CENP-A deposition and maintenance at centromeres.