The fully linked HTML version of this article has now been published.
JCS ePress
online publication date 12 Oct 2004
doi: 10.1242/jcs.01401
Research Article
Elongation of centriolar microtubule triplets contributes to the formation of the mitotic spindle in 
-tubulin-depleted cells
Brigitte Raynaud-Messina*,
Laurent Mazzolini,
André Moisand,
Anne-Marie Cirinesi,
and
Michel Wright
* Author for correspondence (e-mail: brigitte.raynaud-messina{at}istmt.cnrs.fr)
The assembly of the mitotic spindle after depletion of the major 
-tubulin isotype by RNA-mediated interference was assessed in the Drosophila S2 cell line. Depletion of -tubulin had no significant effect on the cytoskeletal microtubules during interphase. However, it promoted an increase in the mitotic index, resulting mainly in monopolar and, to a lesser extent, asymmetrical bipolar prometaphases lacking astral microtubules. This mitotic accumulation coincided with the activation of the mitotic checkpoint. Immunostaining with an anti-Asp antibody revealed that the spindle poles, which were always devoid of -tubulin, were unfocused and organized into sub-spindles. Despite the marked depletion of -tubulin, the pericentriolar proteins CP190 and centrosomin were recruited to the spindle pole(s), where they formed three or four dots, suggesting the presence of several centrioles. Electron microscopic reconstructions demonstrated that most of the monopolar spindles exhibited three or four centrioles, indicating centriole duplication with a failure in the separation process. Most of the centrioles were shortened, suggesting a role for -tubulin in centriole morphogenesis. Moreover, in contrast to metaphases observed in control cells, in which the spindle microtubules radiated from the pericentriolar material, in -tubulin-depleted cells, microtubule assembly still occurred at the poles but involved the elongation of centriolar microtubule triplets. Our results demonstrate that, after depletion of -tubulin, the pericentriolar material is unable to promote efficient microtubule nucleation. They point to an alternative mechanism of centrosomal microtubule assembly that contributes to the formation of abnormal, albeit partially functional, mitotic spindles.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati 
Twitter What's this?
This article has been cited by other articles:
|
|
|
|
 
Y. Xiong and B. R. Oakley
In vivo analysis of the functions of {gamma}-tubulin-complex proteins
J. Cell Sci.,
November 15, 2009;
122(22):
4218 - 4227.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Bouissou, C. Verollet, A. Sousa, P. Sampaio, M. Wright, C. E. Sunkel, A. Merdes, and B. Raynaud-Messina
{gamma}-Tubulin ring complexes regulate microtubule plus end dynamics
J. Cell Biol.,
November 2, 2009;
187(3):
327 - 334.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. Kuriyama, M. Bettencourt-Dias, I. Hoffmann, M. Arnold, and L. Sandvig
{gamma}-Tubulin-containing abnormal centrioles are induced by insufficient Plk4 in human HCT116 colorectal cancer cells
J. Cell Sci.,
June 15, 2009;
122(12):
2014 - 2023.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
X. Fant, N. Gnadt, L. Haren, and A. Merdes
Stability of the small {gamma}-tubulin complex requires HCA66, a protein of the centrosome and the nucleolus
J. Cell Sci.,
April 15, 2009;
122(8):
1134 - 1144.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
G. C. Rogers, N. M. Rusan, M. Peifer, and S. L. Rogers
A Multicomponent Assembly Pathway Contributes to the Formation of Acentrosomal Microtubule Arrays in Interphase Drosophila Cells
Mol. Biol. Cell,
July 1, 2008;
19(7):
3163 - 3178.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Dammermann, P. S. Maddox, A. Desai, and K. Oegema
SAS-4 is recruited to a dynamic structure in newly forming centrioles that is stabilized by the {gamma}-tubulin-mediated addition of centriolar microtubules
J. Cell Biol.,
February 25, 2008;
180(4):
771 - 785.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Wiese
Distinct Dgrip84 Isoforms Correlate with Distinct {gamma}-Tubulins in Drosophila
Mol. Biol. Cell,
January 1, 2008;
19(1):
368 - 377.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. Boutros, V. Lobjois, and B. Ducommun
CDC25B Involvement in the Centrosome Duplication Cycle and in Microtubule Nucleation
Cancer Res.,
December 15, 2007;
67(24):
11557 - 11564.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. S. Taylor, K. G. Hardwick, K. E. Sawin, S. Biggins, S. Piatti, A. Khodjakov, C. L. Rieder, E. D. Salmon, and A. Musacchio
Comment on "A Centrosome-Independent Role for {gamma}-TuRC Proteins in the Spindle Assembly Checkpoint"
Science,
May 18, 2007;
316(5827):
982b - 982b.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. Binarova, V. Cenklova, J. Prochazkova, A. Doskocilova, J. Volc, M. Vrlik, and L. Bogre
{gamma}-Tubulin Is Essential for Acentrosomal Microtubule Nucleation and Coordination of Late Mitotic Events in Arabidopsis
PLANT CELL,
May 1, 2006;
18(5):
1199 - 1212.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. Haren, M.-H. Remy, I. Bazin, I. Callebaut, M. Wright, and A. Merdes
NEDD1-dependent recruitment of the {gamma}-tubulin ring complex to the centrosome is necessary for centriole duplication and spindle assembly
J. Cell Biol.,
February 13, 2006;
172(4):
505 - 515.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Verollet, N. Colombie, T. Daubon, H.-M. Bourbon, M. Wright, and B. Raynaud-Messina
Drosophila melanogaster {gamma}-TuRC is dispensable for targeting {gamma}-tubulin to the centrosome and microtubule nucleation
J. Cell Biol.,
February 13, 2006;
172(4):
517 - 528.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
N. Colombie, C. Verollet, P. Sampaio, A. Moisand, C. Sunkel, H.-M. Bourbon, M. Wright, and B. Raynaud-Messina
The Drosophila {gamma}-Tubulin Small Complex Subunit Dgrip84 Is Required for Structural and Functional Integrity of the Spindle Apparatus
Mol. Biol. Cell,
January 1, 2006;
17(1):
272 - 282.
[Abstract]
[Full Text]
[PDF]
|
|
|
© The Company of Biologists Ltd 2004
