|
|
|
|
|
|
|
|
|||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | |||||
Journal of Cell Science, Vol 108, Issue 3 1083-1092, Copyright © 1995 by Company of Biologists
JOURNAL ARTICLES |
A Vassilev, M Kimble, CD Silflow, M LaVoie and R Kuriyama
Department of Cell Biology and Neuroanatomy, University of Minnesota, Minneapolis 55455, USA.
A new member of the tubulin superfamily, gamma-tubulin, is localized at microtubule-organizing centers (MTOCs) in a variety of organisms. Chlamydomonas cDNA coding for the full-length sequence of gamma-tubulin was expressed in insect ovarian Sf9 cells using the baculovirus expression system. Approximately half of the induced 52 kDa gamma-tubulin was recovered in the supernatant after centrifugation of Sf9 cell lysates at 18,000 g for 15 minutes. When the cell supernatant was analyzed by FPLC on a Superdex 200 sizing column, Chlamydomonas gamma-tubulin separated into two major peaks. The lagging peak contained a monomeric form of gamma-tubulin with a sedimentation coefficient of 2.5 S, which interacted with the Superdex column in a salt-dependent manner. The leading peak, with an apparent molecular mass of 900 kDa, corresponded to a molecular chaperonin complex, and TCP1 chaperonin released folded gamma-tubulin polypeptide from the complex in the presence of MgATP. The released gamma-tubulin monomers were capable of binding to microtubules in vitro and biochemical quantities of active monomers were further purified using a combination of size-exclusion and ion-exchange column chromatography. The endogenous Sf9 cell gamma-tubulin migrated faster than Chlamydomonas gamma-tubulin with an apparent molecular mass of 49 kDa on gels. Analyses on gel filtration and sucrose density gradient centrifugation showed that, while overexpressed Chlamydomonas gamma-tubulin was present in a monomeric form, endogenous gamma-tubulin from Sf9 and HeLa cells exists as a dimer. These results may suggest the possibility that gamma-tubulin could form a heterodimer with hitherto unknown molecule(s).
This article has been cited by other articles:
|
|
 |
|
  S. Fromherz, T. H. Giddings Jr, N. Gomez-Ospina, and S. K. Dutcher Mutations in {alpha}-tubulin promote basal body maturation and flagellar assembly in the absence of {delta}-tubulin J. Cell Sci., January 15, 2004; 117(2): 303 - 314. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. N. Gunawardane, O. C. Martin, and Y. Zheng Characterization of a New gamma TuRC Subunit with WD Repeats Mol. Biol. Cell, March 1, 2003; 14(3): 1017 - 1026. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Drykova, V. Cenklova, V. Sulimenko, J. Volc, P. Draber, and P. Binarova Plant {gamma}-Tubulin Interacts with {alpha}{beta}-Tubulin Dimers and Forms Membrane-Associated Complexes PLANT CELL, February 1, 2003; 15(2): 465 - 480. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Caplow and L. Fee Dissociation of the Tubulin Dimer Is Extremely Slow, Thermodynamically Very Unfavorable, and Reversible in the Absence of an Energy Source Mol. Biol. Cell, June 1, 2002; 13(6): 2120 - 2131. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. K. Dutcher and E. C. Trabuco The UNI3 Gene Is Required for Assembly of Basal Bodies of Chlamydomonas and Encodes delta -Tubulin, a New Member of the Tubulin Superfamily Mol. Biol. Cell, June 1, 1998; 9(6): 1293 - 1308. [Abstract] [Full Text]
|
|
|
|
|
|
A Paoletti, M Moudjou, M Paintrand, J. Salisbury, and M Bornens Most of centrin in animal cells is not centrosome-associated and centrosomal centrin is confined to the distal lumen of centrioles J. Cell Sci., January 12, 1996; 109(13): 3089 - 3102. [Abstract] [PDF]
|
|
|
|
|
|
M Moudjou, N Bordes, M Paintrand, and M Bornens gamma-Tubulin in mammalian cells: the centrosomal and the cytosolic forms J. Cell Sci., January 4, 1996; 109(4): 875 - 887. [Abstract] [PDF]
|
|
|
|
 |
|
 R. Leguy, R. Melki, D. Pantaloni, and M.-F. Carlier Monomeric gamma -Tubulin Nucleates Microtubules J. Biol. Chem., July 14, 2000; 275(29): 21975 - 21980. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Ohta, R. Essner, J.-H. Ryu, R. E. Palazzo, Y. Uetake, and R. Kuriyama Characterization of Cep135, a novel coiled-coil centrosomal protein involved in microtubule organization in mammalian cells J. Cell Biol., January 7, 2002; 156(1): 87 - 100. [Abstract] [Full Text] [PDF]
|
|
