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PMID: 7559777 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

gamma-tubulin is a minus end-specific microtubule binding protein.

The Journal of cell biology ·Vol. 131 ·No. 1 ·1995-10-00 ·Pages 207-14

Li Q, Joshi HC

Abstract

The role of microtubules in mediating chromosome segregation during mitosis is well-recognized. In addition, interphase cells depend upon a radial and uniform orientation of microtubules, which are intrinsically asymmetric polymers, for the directional transport of many cytoplasmic components and for the maintenance of the structural integrity of certain organelles. The slow growing minus ends of microtubules are linked to the centrosome ensuring extension of the fast growing plus ends toward the cell periphery. However, the molecular mechanism of this linkage is not clear. One hypothesis is that gamma-tubulin, located at the centrosome, binds to the minus ends of microtubules. To test this model, we synthesized radiolabeled gamma-tubulin in vitro. We demonstrate here biochemically a specific, saturable, and tight (Kd = 10(-10) M) interaction of gamma-tubulin and microtubule ends with a stoichiometry of 12.6 +/- 4.9 molecules of gamma-tubulin per microtubule. In addition, we designed an in vitro assay to visualize gamma-tubulin at the minus ends of axonemal microtubules. These data show that gamma-tubulin represents the first protein to bind microtubule minus ends and might be responsible for mediating the link between microtubules and the centrosome.

MeSH Terms
Autoradiography Humans Microtubule-Associated Proteins/metabolism Microtubules/metabolism Protein Binding/physiology Recombinant Proteins/metabolism Tubulin/metabolism
Chemicals
Microtubule-Associated Proteins Recombinant Proteins Tubulin
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Li Q
Graduate Program in Biochemistry and Molecular Biology, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
Joshi H C
References (61)
61 references, click to expand
  1. Mitotic spindle organization by a plus-end-directed microtubule motor.
    Nature. 1992 Oct 8;359(6395):540-3 PMID: 1406972
  2. Purification and assay of microtubule-associated proteins (MAPs).
    Methods Enzymol. 1982;85 Pt B:409-16 PMID: 7121279
  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. Rethinking mitosis.
    Cell. 1982 Jul;29(3):729-44 PMID: 6217897
  5. The anatomy of flagellar microtubules: polarity, seam, junctions, and lattice.
    J Cell Biol. 1995 Jan;128(1-2):81-94 PMID: 7822425
  6. 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
  7. Localization of an exchangeable GTP binding site at the plus end of microtubules.
    Science. 1993 Aug 20;261(5124):1044-7 PMID: 8102497
  8. Reversible assembly purification of microtubules without assembly-promoting agents and further purification of tubulin, microtubule-associated proteins, and MAP fragments.
    Methods Enzymol. 1986;134:89-104 PMID: 3821583
  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. The macronuclear gamma-tubulin-encoding gene of Euplotes octocarinatus contains two introns and an in-frame TGA.
    Gene. 1993 Dec 22;136(1-2):319-22 PMID: 8294024
  12. In vitro microtubule-nucleating activity of spindle pole bodies in fission yeast Schizosaccharomyces pombe: cell cycle-dependent activation in xenopus cell-free extracts.
    J Cell Biol. 1992 Jun;117(5):1055-66 PMID: 1533643
  13. 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
  14. Measurement of protein using bicinchoninic acid.
    Anal Biochem. 1985 Oct;150(1):76-85 PMID: 3843705
  15. The gamma-tubulin-encoding gene from the basidiomycete fungus, Ustilago violacea, has a long 5'-untranslated region.
    Gene. 1993 Dec 31;137(2):187-94 PMID: 8299946
  16. gamma-Tubulin is present in acentriolar MTOCs during early mouse development.
    J Cell Sci. 1993 May;105 ( Pt 1):157-66 PMID: 8360270
  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. Isolation, characterization and sequence of a cDNA encoding gamma-tubulin protein from the fern Anemia phyllitidis L. Sw.
    Plant Mol Biol. 1993 Nov;23(3):595-603 PMID: 8219092
  20. 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
  21. Preparation of modified tubulins.
    Methods Enzymol. 1991;196:478-85 PMID: 2034137
  22. Gamma-tubulin is a highly conserved component of the centrosome.
    Cell. 1991 May 31;65(5):825-36 PMID: 1840506
  23. Gamma-tubulin can both nucleate microtubule assembly and self-assemble into novel tubular structures in mammalian cells.
    J Cell Biol. 1995 Sep;130(5):1137-47 PMID: 7657698
  24. Gamma-tubulin is required for the structure and function of the microtubule organizing centre in Drosophila neuroblasts.
    EMBO J. 1995 Jan 3;14(1):28-36 PMID: 7828594
  25. 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
  26. Gamma-tubulin is a centrosomal protein required for cell cycle-dependent microtubule nucleation.
    Nature. 1992 Mar 5;356(6364):80-3 PMID: 1538786
  27. gamma-Tubulin in Arabidopsis: gene sequence, immunoblot, and immunofluorescence studies.
    Plant Cell. 1994 Feb;6(2):303-14 PMID: 8148650
  28. Gamma-tubulin reorganization during mouse fertilization and early development.
    J Cell Sci. 1993 Feb;104 ( Pt 2):383-9 PMID: 8505367
  29. Tubulin domains probed by limited proteolysis and subunit-specific antibodies.
    J Mol Biol. 1985 Sep 20;185(2):311-27 PMID: 4057249
  30. The pericentriolar material in Chinese hamster ovary cells nucleates microtubule formation.
    J Cell Biol. 1977 Jun;73(3):601-15 PMID: 559676
  31. Tublin: nucleotide binding and enzymic activity.
    J Mol Biol. 1974 Nov 5;89(3):455-68 PMID: 4280448
  32. Microtubule assembly and nucleation.
    Int Rev Cytol. 1978;54:1-71 PMID: 391755
  33. Exchange of tubulin dimer into rings in microtubule assembly--disassembly.
    Biochemistry. 1980 Oct 28;19(22):5078-86 PMID: 7459325
  34. A three-domain structure of kinesin heavy chain revealed by DNA sequence and microtubule binding analyses.
    Cell. 1989 Mar 10;56(5):879-89 PMID: 2522352
  35. Isolation of mip (microtubule-interacting protein) mutations of Aspergillus nidulans.
    Mol Cell Biol. 1986 Aug;6(8):2963-8 PMID: 3537728
  36. Localization of the exchangeable nucleotide binding domain in beta-tubulin.
    Biochem Biophys Res Commun. 1986 Mar 28;135(3):1135-43 PMID: 3964271
  37. Microtubule organizing centers.
    Annu Rev Cell Biol. 1985;1:145-72 PMID: 3916316
  38. 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
  39. Structural and functional reconstitution of inner dynein arms in Chlamydomonas flagellar axonemes.
    J Cell Biol. 1992 May;117(3):573-81 PMID: 1533396
  40. Gamma-tubulin distribution in the neuron: implications for the origins of neuritic microtubules.
    J Cell Biol. 1992 Oct;119(1):171-8 PMID: 1527168
  41. Polewards microtubule flux in the mitotic spindle: evidence from photoactivation of fluorescence.
    J Cell Biol. 1989 Aug;109(2):637-52 PMID: 2760109
  42. 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
  43. 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
  44. Determination of free and bound microtubular protein and guanine nucleotide under equilibrium conditions.
    Biochemistry. 1979 Sep 4;18(18):3880-6 PMID: 486401
  45. Interactions of a photoaffinity analog of GTP with the proteins of microtubules.
    Proc Natl Acad Sci U S A. 1977 Oct;74(10):4375-7 PMID: 270679
  46. Role of microtubules in the organisation of the Golgi apparatus.
    Cell Motil Cytoskeleton. 1990;15(2):67-70 PMID: 2178782
  47. Apical orientation of the microtubule organizing center and associated gamma-tubulin during the polarization of the retinal pigment epithelium in vivo.
    Dev Biol. 1993 May;157(1):147-56 PMID: 8482407
  48. Chaperonin-mediated folding of vertebrate actin-related protein and gamma-tubulin.
    J Cell Biol. 1993 Sep;122(6):1301-10 PMID: 8104191
  49. 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
  50. 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
  51. 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
  52. Gamma-tubulin is associated with a cortical-microtubule-organizing zone in the developing guard cells of Allium cepa L.
    Planta. 1993;191(3):357-61 PMID: 7764024
  53. Pericentrin, a highly conserved centrosome protein involved in microtubule organization.
    Cell. 1994 Feb 25;76(4):639-50 PMID: 8124707
  54. Recent progress on structural interactions of the endoplasmic reticulum.
    Cell Motil Cytoskeleton. 1990;15(2):71-5 PMID: 2178783
  55. In vitro reconstitution of centrosome assembly and function: the central role of gamma-tubulin.
    Cell. 1994 Feb 25;76(4):623-37 PMID: 8124706
  56. Gamma-tubulin: the microtubule organizer?
    Trends Cell Biol. 1992 Jan;2(1):1-5 PMID: 14731630
  57. Visualization of the structural polarity of microtubules.
    Nature. 1980 Jul 31;286(5772):517-9 PMID: 7402333
  58. Isolation of Chlamydomonas flagella and flagellar axonemes.
    Methods Enzymol. 1986;134:280-90 PMID: 3821567
  59. 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
  60. 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
  61. 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
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.131.1.207
Published
1995-10-00
Pages
207-14
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2120595
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.131.1.207 PMC Full Text View journal details
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