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Vol. 12, Issue 6, 1751-1764, June 2001
-Tubulin in Early
Caenorhabditis elegans Embryos
and
*Department of Biology, Indiana University, Bloomington,
Indiana 47405-3700; Department of Molecular Biology and
Genetics, Johns Hopkins University, School of Medicine, Baltimore,
Maryland 21205-2185; and §Laboratory of Molecular Biology,
University of Wisconsin, Madison, Wisconsin 53706
-Tubulin is a ubiquitous and highly conserved component of
centrosomes in eukaryotic cells. Genetic and biochemical studies have
demonstrated that -tubulin functions as part of a complex to
nucleate microtubule polymerization from centrosomes. We show that, as
in other organisms, Caenorhabditis elegans -tubulin is concentrated in centrosomes. To study centrosome dynamics in embryos, we generated transgenic worms that express
GFP::-tubulin or GFP::
-tubulin in the
maternal germ line and early embryos. Multiphoton microscopy of embryos
produced by these worms revealed the time course of daughter centrosome
appearance and growth and the differential behavior of centrosomes
destined for germ line and somatic blastomeres. To study the role of
-tubulin in nucleation and organization of spindle microtubules, we
used RNA interference (RNAi) to deplete C. elegans
embryos of -tubulin. -Tubulin (RNAi) embryos failed in
chromosome segregation, but surprisingly, they contained extensive
microtubule arrays. Moderately affected embryos contained bipolar
spindles with dense and long astral microtubule arrays but with poorly
organized kinetochore and interpolar microtubules. Severely
affected embryos contained collapsed spindles with numerous long astral
microtubules. Our results suggest that -tubulin is not absolutely
required for microtubule nucleation in C. elegans but is
required for the normal organization and function of
kinetochore and interpolar microtubules.
Online version of this article contains video material
for Figures 4, 5, 8, and 9. Online version available at
www.molbiolcell.org.
Corresponding author. E-mail
address: sstrome{at}bio.indiana.edu.
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