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Vol. 11, Issue 6, 2047-2056, June 2000
Tubulin onto Centrosomes
and
*Program in Molecular Medicine and Centrosome assembly is important for mitotic spindle formation and
if defective may contribute to genomic instability in cancer. Here we
show that in somatic cells centrosome assembly of two proteins involved
in microtubule nucleation, pericentrin and Biomedical Imaging
Group, University of Massachusetts Medical School, Worcester,
Massachusetts 01605
tubulin, is inhibited in
the absence of microtubules. A more potent inhibitory effect on
centrosome assembly of these proteins is observed after specific
disruption of the microtubule motor cytoplasmic dynein by
microinjection of dynein antibodies or by overexpression of the
dynamitin subunit of the dynein binding complex dynactin. Consistent
with these observations is the ability of pericentrin to cosediment
with taxol-stabilized microtubules in a dynein- and dynactin-dependent
manner. Centrosomes in cells with reduced levels of pericentrin and tubulin have a diminished capacity to nucleate microtubules. In living
cells expressing a green fluorescent protein-pericentrin fusion
protein, green fluorescent protein particles containing endogenous
pericentrin and tubulin move along microtubules at speeds of dynein
and dock at centrosomes. In Xenopus extracts where tubulin assembly onto centrioles can occur without microtubules, we
find that assembly is enhanced in the presence of microtubules and
inhibited by dynein antibodies. From these studies we conclude that
pericentrin and tubulin are novel dynein cargoes that can be
transported to centrosomes on microtubules and whose assembly
contributes to microtubule nucleation.
Online version of this article contains video
material for fig. 1. Online version available at www.molbiolcell.org.
Corresponding author. E-mail address:
stephen.doxsey{at}ummed.edu.
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S. G. Addinall, P. S. Mayr, S. Doyle, J. K. Sheehan, P. G. Woodman, and V. J. Allan Phosphorylation by cdc2-CyclinB1 Kinase Releases Cytoplasmic Dynein from Membranes J. Biol. Chem., May 4, 2001; 276(19): 15939 - 15944. [Abstract] [Full Text] [PDF]
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P. S. Vaughan, J. D. Leszyk, and K. T. Vaughan Cytoplasmic Dynein Intermediate Chain Phosphorylation Regulates Binding to Dynactin J. Biol. Chem., July 6, 2001; 276(28): 26171 - 26179. [Abstract] [Full Text] [PDF]
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I. Vorobjev, V. Malikov, and V. Rodionov Self-organization of a radial microtubule array by dynein-dependent nucleation of microtubules PNAS, August 28, 2001; 98(18): 10160 - 10165. [Abstract] [Full Text] [PDF]
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