title: Assembly of Drosophila centromeric chromatin proteins during mitosis.
abstract: Semi-conservative segregation of nucleosomes to sister chromatids during DNA replication creates gaps that must be filled by new nucleosome assembly. We analyzed the cell-cycle timing of centromeric chromatin assembly in Drosophila, which contains the H3 variant CID (CENP-A in humans), as well as CENP-C and CAL1, which are required for CID localization. Pulse-chase experiments show that CID and CENP-C levels decrease by 50% at each cell division, as predicted for semi-conservative segregation and inheritance, whereas CAL1 displays higher turnover. Quench-chase-pulse experiments demonstrate that there is a significant lag between replication and replenishment of centromeric chromatin. Surprisingly, new CID is recruited to centromeres in metaphase, by a mechanism that does not require an intact mitotic spindle, but does require proteasome activity. Interestingly, new CAL1 is recruited to centromeres before CID in prophase. Furthermore, CAL1, but not CENP-C, is found in complex with pre-nucleosomal CID. Finally, CENP-C displays yet a different pattern of incorporation, during both interphase and mitosis. The unusual timing of CID recruitment and unique dynamics of CAL1 identify a distinct centromere assembly pathway in Drosophila and suggest that CAL1 is a key regulator of centromere propagation.
authors: Department of Molecular and Cell Biology, University of Connecticut, Storrs, Connecticut, United States of America. barbara.mellone@uconn.edu
authors: Barbara G
authors: BG
authors: Mellone
authors: Kathryn J
authors: KJ
authors: Grive
authors: Vladimir
authors: V
authors: Shteyn
authors: Sarion R
authors: SR
authors: Bowers
authors: Isaac
authors: I
authors: Oderberg
authors: Gary H
authors: GH
authors: Karpen
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