HCP-4, a CENP-C-like Protein in Caenorhabditis elegans, Is Required for Resolution of Sister Centromeres

doi:10.1083/jcb.153.6.1199

Published online 4 June 2001. doi:10.1083/jcb.153.6.1199

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© The Rockefeller University Press, 0021-9525/2001/6/1199/ $5.00
The Journal of Cell Biology, Volume 153, Number 6, June 11, 2001 1199-1208

Original Article

HCP-4, a CENP-C–like Protein in Caenorhabditis elegans, Is Required for Resolution of Sister Centromeres

Landon L. Moore^a and Mark B. Roth^a
^a Division of Basic Sciences, Molecular and Cellular Biology Program, Fred Hutchinson Cancer Research Center, Seattle, Washington 98109

Correspondence to: Mark B. Roth, Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue North, Mailstop A3-013, Seattle, WA 98109. Tel:(206) 667-5602 Fax:(206) 667-6877 E-mail:mroth{at}fred.fhcrc.org.

The centromere plays a critical role in the segregation of chromosomesduring mitosis. In mammals, sister centromeres are resolvedfrom one another in the G2 phase of the cell cycle. During prophase,chromosomes condense with sister centromeres oriented in a backto back configuration enabling only one chromatid to be capturedby each half spindle. To study this process, we identified acentromere protein (CENP)-C–like protein, holocentricprotein (HCP)-4, in Caenorhabditis elegans based on sequenceidentity, loss of function phenotype, and centromeric localization.HCP-4 is found in the cytoplasm during interphase, but is nuclearlocalized in mitosis, where it localizes specifically to thecentromere. The localization of HCP-4 to the centromere is dependenton the centromeric histone HCP-3; in addition, HCP-3 and HCP-4are both required for localization of a CENP-F–like protein,HCP-1, indicating an ordered assembly pathway. Loss of HCP-4expression by RNA-mediated interference resulted in a failureto 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 bothresolution sister centromeres and attachment to the mitoticspindle.

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

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