Analysis of kinetochore assembly and function in Caenorhabditis elegans embryos and human cells.
All eukaryotes rely on multi-protein assemblies, called kinetochores, to direct the segregation of their chromosomes in mitosis. The list of known kinetochore components has been growing rapidly in the post-genomic era: in animal cells, there are presently more than 80 proteins that show either exclusive or partial localization at kinetochores during mitosis. The future challenge is to elucidate how these proteins contribute to kinetochore structure, spindle microtubule attachment, regulation of microtubule dynamics, and the detection, signaling, and correction of microtubule attachment errors. Cultured human tumor cells, especially HeLa cells, are widely used for the study of kinetochores. Recently, the experimental advantages offered by the nematode Caenorhabditis elegans have been exploited for functional analysis of kinetochore components in the first embryonic division. Here, we discuss basic methods, largely based on fluorescence imaging, to study kinetochore structure and function in these two metazoan model systems.
Gassmann
Reto
R
Ludwig Institute for Cancer Research, Department of Cellular and Molecular Medicine, UCSD School of Medicine, CMM-East, Rm 3080, 9500 Gilman Dr., La Jolla, CA 92093, USA. rgassmann@ucsd.edu
Kline
Susan L
SL
Carvalho
Ana
A
Desai
Arshad
A
D000818:Animals; D017173:Caenorhabditis elegans / Q000166:cytology / Q000196:embryology / Q000502:physiology*; D045744:Cell Line, Tumor; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D018386:Kinetochores / Q000502:physiology* / Q000648:ultrastructure*; D008856:Microscopy, Fluorescence