Home LiteratureArticle Details
PMID: 7657698 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Gamma-tubulin can both nucleate microtubule assembly and self-assemble into novel tubular structures in mammalian cells.

The Journal of cell biology ·Vol. 130 ·No. 5 ·1995-09-00 ·Pages 1137-47

Shu HB, Joshi HC

Abstract

alpha-, beta-, and gamma-tubulins are evolutionarily highly conserved members of the tubulin gene superfamily. While the abundant members, alpha- and beta-tubulins, constitute the building blocks of cellular microtubule polymers, gamma-tubulin is a low abundance protein which localized to the pericentriolar material and may play a role in microtubule assembly. To test whether gamma-tubulin mediates the nucleation of microtubule assembly in vivo, and co-assembles with alpha- and beta-tubulins into microtubules or self-assembles into macro-molecular structures, we experimentally elevated the expression of gamma-tubulin in the cell cytoplasm. In most cells, overexpression of gamma-tubulin causes a dramatic reorganization of the cellular microtubule network. Furthermore, we show that when overexpressed, gamma-tubulin causes ectopic nucleation of microtubules which are not associated with the centrosome. In a fraction of cells, gamma-tubulin self-assembles into novel tubular structures with a diameter of approximately 50 nm (named gamma-tubules). Furthermore, unlike microtubules, gamma-tubules are resistant to cold or drug induced depolymerization. These data provide evidence that gamma-tubulin can cause nucleation of microtubule assembly and can self-assemble into novel tubular structures.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Cells, Cultured/cytology Centrosome/physiology Cytoplasm/physiology Fluorescent Antibody Technique Gene Expression/physiology Haplorhini Immunohistochemistry Microscopy, Electron Microtubules/physiology,ultrastructure Molecular Sequence Data Nocodazole/pharmacology Polymers/metabolism Rabbits Thermodynamics Tubulin/genetics,metabolism,pharmacology,physiology
Chemicals
Polymers Tubulin Nocodazole
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Shu H B
Department of Anatomy and Cell Biology, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
Joshi H C
References (60)
60 references, click to expand
  1. In vivo microtubules are copolymers of available beta-tubulin isotypes: localization of each of six vertebrate beta-tubulin isotypes using polyclonal antibodies elicited by synthetic peptide antigens.
    J Cell Biol. 1987 Oct;105(4):1707-20 PMID: 3312237
  2. Microtubule-macrotubule transformations induced by volatile anesthetics. Mechanism of macrotubule assembly.
    J Ultrastruct Res. 1976 Dec;57(3):237-50 PMID: 63568
  3. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  4. Gamma-tubulin: the hub of cellular microtubule assemblies.
    Bioessays. 1993 Oct;15(10):637-43 PMID: 8274140
  5. Gamma-tubulin is present in Drosophila melanogaster and Homo sapiens and is associated with the centrosome.
    Cell. 1991 May 31;65(5):817-23 PMID: 1904010
  6. Interpolar spindle microtubules in PTK cells.
    J Cell Biol. 1993 Dec;123(6 Pt 1):1475-89 PMID: 8253845
  7. Vinblastine-induced paracrystals and unusually large microtubules (macrotubules) in rat renal cells.
    Z Zellforsch Mikrosk Anat. 1973 Aug 14;141(4):443-58 PMID: 4355873
  8. Alpha-, beta-, and gamma-tubulins: sequence comparisons and structural constraints.
    Cell Motil Cytoskeleton. 1991;20(3):181-9 PMID: 1773446
  9. Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product.
    Mol Cell Biol. 1985 Dec;5(12):3610-6 PMID: 3915782
  10. Initiation and growth of microtubules from mitotic centers in lysed mammalian cells.
    J Cell Biol. 1975 Dec;67(3):744-60 PMID: 1202022
  11. Identification with cellular microtubules of one of the co-assemlbing microtubule-associated proteins.
    Cell. 1979 Oct;18(2):431-8 PMID: 498277
  12. Identification of gamma-tubulin, a new member of the tubulin superfamily encoded by mipA gene of Aspergillus nidulans.
    Nature. 1989 Apr 20;338(6217):662-4 PMID: 2649796
  13. Measurement of protein using bicinchoninic acid.
    Anal Biochem. 1985 Oct;150(1):76-85 PMID: 3843705
  14. Human gamma-tubulin functions in fission yeast.
    J Cell Biol. 1994 Sep;126(6):1465-73 PMID: 8089179
  15. Immunogold electron microscopy: mapping tubulin isotypes on neurite microtubules.
    Methods Cell Biol. 1993;37:259-81 PMID: 7504779
  16. Dynamic instability of microtubule growth.
    Nature. 1984 Nov 15-21;312(5991):237-42 PMID: 6504138
  17. In vitro polymerization of microtubules into asters and spindles in homogenates of surf clam eggs.
    J Cell Biol. 1975 Jan;64(1):146-58 PMID: 45844
  18. Microtubule nucleating sites in higher plant cells identified by an auto-antibody against pericentriolar material.
    J Cell Biol. 1985 Jul;101(1):319-24 PMID: 4008533
  19. In vitro assembly of multiprotein complexes containing alpha, beta, and gamma tubulin, heat shock protein HSP70, and elongation factor 1 alpha.
    Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):3028-32 PMID: 8464918
  20. Gamma-tubulin is a highly conserved component of the centrosome.
    Cell. 1991 May 31;65(5):825-36 PMID: 1840506
  21. Electron-microscopic study of the spindle and chromosome movement in the yeast Saccharomyces cerevisiae.
    J Cell Sci. 1976 Nov;22(2):219-42 PMID: 794073
  22. Localization of the centrin-related 165,000-Mr protein of PtK2 cells during the cell cycle.
    Cell Motil Cytoskeleton. 1991;18(1):1-14 PMID: 2004430
  23. Gamma-tubulin reorganization during mouse fertilization and early development.
    J Cell Sci. 1993 Feb;104 ( Pt 2):383-9 PMID: 8505367
  24. The Rous sarcoma virus long terminal repeat is a strong promoter when introduced into a variety of eukaryotic cells by DNA-mediated transfection.
    Proc Natl Acad Sci U S A. 1982 Nov;79(22):6777-81 PMID: 6294651
  25. The pericentriolar material in Chinese hamster ovary cells nucleates microtubule formation.
    J Cell Biol. 1977 Jun;73(3):601-15 PMID: 559676
  26. Microtubule assembly and nucleation.
    Int Rev Cytol. 1978;54:1-71 PMID: 391755
  27. Purification and characterization of oocyte cytoplasmic tubulin and meiotic spindle tubulin of the surf clam Spisula solidissima.
    J Cell Biol. 1984 Jan;98(1):253-66 PMID: 6538572
  28. Microtubule organizing centers.
    Annu Rev Cell Biol. 1985;1:145-72 PMID: 3916316
  29. Centrosome assembly in vitro: role of gamma-tubulin recruitment in Xenopus sperm aster formation.
    J Cell Biol. 1994 Jan;124(1-2):19-31 PMID: 8294501
  30. Cytoplasmic microtubules in tissue culture cells appear to grow from an organizing structure towards the plasma membrane.
    Proc Natl Acad Sci U S A. 1976 Mar;73(3):867-71 PMID: 1062799
  31. Heterogeneity of tubulin subunits.
    Proc Natl Acad Sci U S A. 1971 Sep;68(9):2028-31 PMID: 5289362
  32. Gamma-tubulin distribution in the neuron: implications for the origins of neuritic microtubules.
    J Cell Biol. 1992 Oct;119(1):171-8 PMID: 1527168
  33. Microtubules: evidence for 13 protofilaments.
    J Cell Biol. 1973 Nov;59(2 Pt 1):267-75 PMID: 4805001
  34. The fission yeast gamma-tubulin is essential for mitosis and is localized at microtubule organizing centers.
    J Cell Sci. 1991 Aug;99 ( Pt 4):693-700 PMID: 1770000
  35. Delineation by lanthanum staining of filamentous elements associated with the surfaces of axonal microtubules.
    J Cell Sci. 1973 Mar;12(2):567-83 PMID: 4122255
  36. Induction of a sheet polymer of tubulin by Zn2+.
    Exp Cell Res. 1976 Jun;100(1):104-10 PMID: 945172
  37. The microtubule-dependent formation of a tubulovesicular network with characteristics of the ER from cultured cell extracts.
    Cell. 1988 Jul 1;54(1):27-35 PMID: 3289756
  38. Studies on the microtubules in heliozoa. II. The effect of low temperature on these structures in the formation and maintenance of the axopodia.
    J Cell Biol. 1967 Jul;34(1):327-43 PMID: 6033539
  39. Expression of NF-L in both neuronal and nonneuronal cells of transgenic mice: increased neurofilament density in axons without affecting caliber.
    J Cell Biol. 1990 Oct;111(4):1543-57 PMID: 2120242
  40. Role of microtubules in the organisation of the Golgi apparatus.
    Cell Motil Cytoskeleton. 1990;15(2):67-70 PMID: 2178782
  41. Tubulin assembly sites and the organization of cytoplasmic microtubules in cultured mammalian cells.
    J Cell Biol. 1981 Sep;90(3):554-62 PMID: 7026576
  42. Differential utilization of beta-tubulin isotypes in differentiating neurites.
    J Cell Biol. 1989 Aug;109(2):663-73 PMID: 2503525
  43. Microtubule organizing centers and gamma-tubulin.
    Curr Opin Cell Biol. 1994 Feb;6(1):54-62 PMID: 8167026
  44. Chaperonin-mediated folding of vertebrate actin-related protein and gamma-tubulin.
    J Cell Biol. 1993 Sep;122(6):1301-10 PMID: 8104191
  45. Human chromosomes and centrioles as nucleating sites for the in vitro assembly of microtubules from bovine brain tubulin.
    J Cell Biol. 1975 Oct;67(1):189-99 PMID: 809450
  46. Drosophila gamma-tubulin is part of a complex containing two previously identified centrosomal MAPs.
    J Cell Biol. 1993 May;121(4):823-35 PMID: 8491775
  47. A gamma-tubulin-related protein associated with the microtubule arrays of higher plants in a cell cycle-dependent manner.
    J Cell Sci. 1993 Apr;104 ( Pt 4):1217-28 PMID: 7686171
  48. In vivo coassembly of a divergent beta-tubulin subunit (c beta 6) into microtubules of different function.
    J Cell Biol. 1987 Nov;105(5):2179-90 PMID: 3316249
  49. Pericentrin, a highly conserved centrosome protein involved in microtubule organization.
    Cell. 1994 Feb 25;76(4):639-50 PMID: 8124707
  50. Recent progress on structural interactions of the endoplasmic reticulum.
    Cell Motil Cytoskeleton. 1990;15(2):71-5 PMID: 2178783
  51. In vitro reconstitution of centrosome assembly and function: the central role of gamma-tubulin.
    Cell. 1994 Feb 25;76(4):623-37 PMID: 8124706
  52. Gamma-tubulin: the microtubule organizer?
    Trends Cell Biol. 1992 Jan;2(1):1-5 PMID: 14731630
  53. Visualization of the structural polarity of microtubules.
    Nature. 1980 Jul 31;286(5772):517-9 PMID: 7402333
  54. Polarity of some motility-related microtubules.
    Proc Natl Acad Sci U S A. 1981 Jan;78(1):372-6 PMID: 6941252
  55. Differential regulation of vascular cell adhesion molecule 1 gene expression by specific NF-kappa B subunits in endothelial and epithelial cells.
    Mol Cell Biol. 1993 Oct;13(10):6283-9 PMID: 7692229
  56. Gamma-tubulin in differentiated cell types: localization in the vicinity of basal bodies in retinal photoreceptors and ciliated epithelia.
    J Cell Sci. 1993 Apr;104 ( Pt 4):1229-37 PMID: 7686172
  57. A new technique for the assay of infectivity of human adenovirus 5 DNA.
    Virology. 1973 Apr;52(2):456-67 PMID: 4705382
  58. The role of the centriolar region in animal cell mitosis. A laser microbeam study.
    J Cell Biol. 1977 Feb;72(2):351-67 PMID: 556728
  59. Gamma-tubulin is a component of the spindle pole body that is essential for microtubule function in Aspergillus nidulans.
    Cell. 1990 Jun 29;61(7):1289-301 PMID: 2194669
  60. Are cytoplasmic microtubules heteropolymers?
    Proc Natl Acad Sci U S A. 1971 Aug;68(8):1762-6 PMID: 5288762
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.130.5.1137
Published
1995-09-00
Pages
1137-47
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2120553
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.130.5.1137 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2