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Published online 11 June 2001. doi:10.1083/jcb.153.6.1199
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© The Rockefeller University Press, 0021-9525/2001//1199 $5.00
The Journal of Cell Biology, Volume 153, Number 6, , 2001 1199-1208

Original Article

Hcp-4, a Cenp-C–Like Protein inCaenorhabditis elegans, Is Required for Resolution of Sister Centromeres



Landon L. Moorea and Mark B. Rotha

a Division of Basic Sciences, Molecular and Cellular Biology Program, Fred Hutchinson Cancer Research Center, Seattle, Washington 98109
Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue North, Mailstop A3-013, Seattle, WA 98109.(206) 667-6877(206) 667-5602

mroth{at}fred.fhcrc.org

The centromere plays a critical role in the segregation of chromosomes during mitosis. In mammals, sister centromeres are resolved from one another in the G2 phase of the cell cycle. During prophase, chromosomes condense with sister centromeres oriented in a back to back configuration enabling only one chromatid to be captured by each half spindle. To study this process, we identified a centromere protein (CENP)-C–like protein, holocentric protein (HCP)-4, in Caenorhabditis elegans based on sequence identity, loss of function phenotype, and centromeric localization. HCP-4 is found in the cytoplasm during interphase, but is nuclear localized in mitosis, where it localizes specifically to the centromere. The localization of HCP-4 to the centromere is dependent on the centromeric histone HCP-3; in addition, HCP-3 and HCP-4 are both required for localization of a CENP-F–like protein, HCP-1, indicating an ordered assembly pathway. Loss of HCP-4 expression by RNA-mediated interference resulted in a failure to generate resolution of sister centromeres on chromosomes, suggesting that HCP-4 is required for sister centromere resolution. These chromosomes also failed to form a functional kinetochore. Thus, the CENP-C–like protein HCP-4 is essential for both resolution sister centromeres and attachment to the mitotic spindle.

Key Words: Caenorhabditis elegans • chromosome • mitosis • centromere • kinetochore

© 2001 The Rockefeller University Press

Abbreviations used in this paper: CENP, centromere protein; HCP, holocentric protein; RNAi, RNA-mediated interference.


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