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PMID: 1348005 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Yeast proteins associated with microtubules in vitro and in vivo.

Molecular biology of the cell ·Vol. 3 ·No. 1 ·1992-01-00 ·Pages 29-47

Barnes G, Louie KA, Botstein D

Abstract

Conditions were established for the self-assembly of milligram amounts of purified Saccharomyces cerevisiae tubulin. Microtubules assembled with pure yeast tubulin were not stabilized by taxol; hybrid microtubules containing substoichiometric amounts of bovine tubulin were stabilized. Yeast microtubule-associated proteins (MAPs) were identified on affinity matrices made from hybrid and all-bovine microtubules. About 25 yeast MAPs were isolated. The amino-terminal sequences of several of these were determined: three were known metabolic enzymes, two were GTP-binding proteins (including the product of the SAR1 gene), and three were novel proteins not found in sequence databases. Affinity-purified antisera were generated against synthetic peptides corresponding to two of the apparently novel proteins (38 and 50 kDa). Immunofluorescence microscopy showed that both these proteins colocalize with intra- and extranuclear microtubules in vivo.

MeSH Terms
Alkaloids/pharmacology Chromatography, Affinity Electrophoresis, Polyacrylamide Gel Fluorescent Antibody Technique Fungal Proteins/chemistry,isolation & purification,metabolism GTP-Binding Proteins/chemistry,isolation & purification,metabolism Microtubule-Associated Proteins/chemistry,isolation & purification,metabolism Microtubules/chemistry Molecular Weight Paclitaxel Saccharomyces cerevisiae/chemistry Tubulin/chemistry,isolation & purification
Chemicals
Alkaloids Fungal Proteins Microtubule-Associated Proteins Tubulin GTP-Binding Proteins Paclitaxel
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Barnes G
Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.
Louie K A
Botstein D
References (64)
64 references, click to expand
  1. Structural rearrangements of tubulin and actin during the cell cycle of the yeast Saccharomyces.
    J Cell Biol. 1984 Mar;98(3):922-33 PMID: 6365930
  2. The cytoskeleton of Saccharomyces cerevisiae.
    Curr Opin Cell Biol. 1990 Feb;2(1):109-15 PMID: 2183834
  3. Microtubule assembly nucleated by isolated centrosomes.
    Nature. 1984 Nov 15-21;312(5991):232-7 PMID: 6504137
  4. Rat monoclonal antitubulin antibodies derived by using a new nonsecreting rat cell line.
    J Cell Biol. 1982 Jun;93(3):576-82 PMID: 6811596
  5. Purification of yeast tubulin by self-assembly in vitro.
    Biochemistry. 1981 Jun 9;20(12):3629-33 PMID: 7020758
  6. High resolution two-dimensional electrophoresis of proteins.
    J Biol Chem. 1975 May 25;250(10):4007-21 PMID: 236308
  7. A mutant of Saccharomyces cerevisiae defective for nuclear fusion.
    Proc Natl Acad Sci U S A. 1976 Oct;73(10):3651-5 PMID: 790391
  8. Multiple nucleotide-binding sites in the sequence of dynein beta heavy chain.
    Nature. 1991 Aug 15;352(6336):640-3 PMID: 1830927
  9. The mitotic apparatus-associated 51-kDa protein from sea urchin eggs is a GTP-binding protein and is immunologically related to yeast polypeptide elongation factor 1 alpha.
    J Biol Chem. 1990 Feb 25;265(6):3240-7 PMID: 2105945
  10. Molecular cloning of the microtubule-associated mechanochemical enzyme dynamin reveals homology with a new family of GTP-binding proteins.
    Nature. 1990 Sep 20;347(6290):256-61 PMID: 2144893
  11. Associations between microtubules and intracellular organelles.
    Curr Opin Cell Biol. 1990 Feb;2(1):105-8 PMID: 2183833
  12. BIK1, a protein required for microtubule function during mating and mitosis in Saccharomyces cerevisiae, colocalizes with tubulin.
    J Cell Biol. 1990 Dec;111(6 Pt 1):2573-86 PMID: 2277073
  13. Fluorescence microscopy methods for yeast.
    Methods Cell Biol. 1989;31:357-435 PMID: 2476649
  14. A novel GTP-binding protein, Sar1p, is involved in transport from the endoplasmic reticulum to the Golgi apparatus.
    J Cell Biol. 1989 Dec;109(6 Pt 1):2677-91 PMID: 2512296
  15. KAR2, a karyogamy gene, is the yeast homolog of the mammalian BiP/GRP78 gene.
    Cell. 1989 Jun 30;57(7):1211-21 PMID: 2661018
  16. Identifying mutations in duplicated functions in Saccharomyces cerevisiae: recessive mutations in HMG-CoA reductase genes.
    Genetics. 1987 Dec;117(4):645-55 PMID: 2828155
  17. Isolation and characterization of mutations in the beta-tubulin gene of Saccharomyces cerevisiae.
    Genetics. 1985 Dec;111(4):715-34 PMID: 2998923
  18. Chromosome instability mutants of Saccharomyces cerevisiae that are defective in microtubule-mediated processes.
    Mol Cell Biol. 1990 Jan;10(1):223-34 PMID: 2403635
  19. Yeast mutants sensitive to antimicrotubule drugs define three genes that affect microtubule function.
    Genetics. 1990 Feb;124(2):251-62 PMID: 2407611
  20. The microtubule binding domain of microtubule-associated protein MAP1B contains a repeated sequence motif unrelated to that of MAP2 and tau.
    J Cell Biol. 1989 Dec;109(6 Pt 2):3367-76 PMID: 2480963
  21. 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
  22. Identification of microtubule-associated proteins in the centrosome, spindle, and kinetochore of the early Drosophila embryo.
    J Cell Biol. 1989 Dec;109(6 Pt 1):2977-91 PMID: 2574177
  23. The pentafunctional arom enzyme of Saccharomyces cerevisiae is a mosaic of monofunctional domains.
    Biochem J. 1987 Sep 1;246(2):375-86 PMID: 2825635
  24. Purification of microtubules and microtubule-associated proteins from sea urchin eggs and cultured mammalian cells using taxol, and use of exogenous taxol-stabilized brain microtubules for purifying microtubule-associated proteins.
    Methods Enzymol. 1986;134:116-27 PMID: 2881189
  25. Isolation and characterization of yeast strains carrying mutations in the glyceraldehyde-3-phosphate dehydrogenase genes.
    J Biol Chem. 1985 Dec 5;260(28):15013-8 PMID: 2999100
  26. Two functional alpha-tubulin genes of the yeast Saccharomyces cerevisiae encode divergent proteins.
    Mol Cell Biol. 1986 Nov;6(11):3711-21 PMID: 3025610
  27. Secretion of invertase in mitotic yeast cells.
    EMBO J. 1988 May;7(5):1475-82 PMID: 3044781
  28. Yeast actin-binding proteins: evidence for a role in morphogenesis.
    J Cell Biol. 1988 Dec;107(6 Pt 2):2551-61 PMID: 3060468
  29. Isolation and characterization of conditional-lethal mutations in the TUB1 alpha-tubulin gene of the yeast Saccharomyces cerevisiae.
    Genetics. 1988 Nov;120(3):681-95 PMID: 3066684
  30. The yeast GTP-binding YPT1 protein and a mammalian counterpart are associated with the secretion machinery.
    Cell. 1988 Mar 25;52(6):915-24 PMID: 3127057
  31. Duck lens epsilon-crystallin and lactate dehydrogenase B4 are identical: a single-copy gene product with two distinct functions.
    Proc Natl Acad Sci U S A. 1988 Oct;85(19):7114-8 PMID: 3174623
  32. Unlinked noncomplementation: isolation of new conditional-lethal mutations in each of the tubulin genes of Saccharomyces cerevisiae.
    Genetics. 1988 Jun;119(2):249-60 PMID: 3294100
  33. Specific binding of [alpha-32P]GTP to cytosolic and membrane-bound proteins of human platelets correlates with the activation of phospholipase C.
    Proc Natl Acad Sci U S A. 1987 Apr;84(8):2261-5 PMID: 3470789
  34. Mitochondrial and nonmitochondrial citrate synthases in Saccharomyces cerevisiae are encoded by distinct homologous genes.
    Mol Cell Biol. 1986 Dec;6(12):4509-15 PMID: 3540614
  35. Microtubule-associated proteins.
    Annu Rev Cell Biol. 1986;2:421-57 PMID: 3548773
  36. Recruitment of enzymes as lens structural proteins.
    Science. 1987 Jun 19;236(4808):1554-6 PMID: 3589669
  37. Differential expression of the three yeast glyceraldehyde-3-phosphate dehydrogenase genes.
    J Biol Chem. 1985 Dec 5;260(28):15019-27 PMID: 3905788
  38. Influence of the centrosome on the structure of nucleated microtubules.
    J Cell Biol. 1985 Apr;100(4):1185-91 PMID: 4038981
  39. Relationships between the structures of taxol and baccatine III derivatives and their in vitro action on the disassembly of mammalian brain and Physarum amoebal microtubules.
    Proc Natl Acad Sci U S A. 1984 Jul;81(13):4090-4 PMID: 6146136
  40. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A.
    Anal Biochem. 1981 Apr;112(2):195-203 PMID: 6266278
  41. KAR1, a gene required for function of both intranuclear and extranuclear microtubules in yeast.
    Cell. 1987 Mar 27;48(6):1047-60 PMID: 3030557
  42. Functions of microtubules in the Saccharomyces cerevisiae cell cycle.
    J Cell Biol. 1988 Oct;107(4):1409-26 PMID: 3049620
  43. The primary structure and heterogeneity of tau protein from mouse brain.
    Science. 1988 Jan 15;239(4837):285-8 PMID: 3122323
  44. Microtubule-associated protein MAP2 shares a microtubule binding motif with tau protein.
    Science. 1988 Nov 11;242(4880):936-9 PMID: 3142041
  45. Diverse effects of beta-tubulin mutations on microtubule formation and function.
    J Cell Biol. 1988 Jun;106(6):1997-2010 PMID: 3290223
  46. Genetically essential and nonessential alpha-tubulin genes specify functionally interchangeable proteins.
    Mol Cell Biol. 1986 Nov;6(11):3722-33 PMID: 3540600
  47. Dynamic and stable populations of microtubules in cells.
    J Cell Biol. 1987 Feb;104(2):277-88 PMID: 3543024
  48. A chicken-yeast chimeric beta-tubulin protein is incorporated into mouse microtubules in vivo.
    Cell. 1986 Feb 14;44(3):461-8 PMID: 3753663
  49. Bundling of microtubules by glyceraldehyde-3-phosphate dehydrogenase and its modulation by ATP.
    Eur J Biochem. 1985 Jul 15;150(2):265-9 PMID: 4018083
  50. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  51. Genetic analysis of yeast RAS1 and RAS2 genes.
    Cell. 1984 Jun;37(2):437-45 PMID: 6327067
  52. Relationship of actin and tubulin distribution to bud growth in wild-type and morphogenetic-mutant Saccharomyces cerevisiae.
    J Cell Biol. 1984 Mar;98(3):934-45 PMID: 6365931
  53. Dynamic instability of microtubule growth.
    Nature. 1984 Nov 15-21;312(5991):237-42 PMID: 6504138
  54. A porcine brain protein (35 K protein) which bundles microtubules and its identification as glyceraldehyde 3-phosphate dehydrogenase.
    J Biochem. 1983 May;93(5):1259-69 PMID: 6885722
  55. Protein measurement with the Folin phenol reagent.
    J Biol Chem. 1951 Nov;193(1):265-75 PMID: 14907713
  56. Promotion of microtubule assembly in vitro by taxol.
    Nature. 1979 Feb 22;277(5698):665-7 PMID: 423966
  57. The three-dimensional structure of tubulin protofilaments.
    Nature. 1979 Jun 14;279(5714):607-12 PMID: 450110
  58. A simplification of the protein assay method of Lowry et al. which is more generally applicable.
    Anal Biochem. 1977 Dec;83(2):346-56 PMID: 603028
  59. Ability of the c-mos product to associate with and phosphorylate tubulin.
    Science. 1991 Feb 8;251(4994):671-5 PMID: 1825142
  60. Four ATP-binding sites in the midregion of the beta heavy chain of dynein.
    Nature. 1991 Aug 15;352(6336):643-5 PMID: 1830928
  61. ADP ribosylation factor is an essential protein in Saccharomyces cerevisiae and is encoded by two genes.
    Mol Cell Biol. 1990 Dec;10(12):6690-9 PMID: 2123295
  62. KAR3, a kinesin-related gene required for yeast nuclear fusion.
    Cell. 1990 Mar 23;60(6):1029-41 PMID: 2138512
  63. Microtubule-dependent retrograde transport of proteins into the ER in the presence of brefeldin A suggests an ER recycling pathway.
    Cell. 1990 Mar 9;60(5):821-36 PMID: 2178778
  64. Isolation of the beta-tubulin gene from yeast and demonstration of its essential function in vivo.
    Cell. 1983 May;33(1):211-9 PMID: 6380751
Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.3.1.29
Published
1992-01-00
Pages
29-47
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC275500
Subset
IM
External Links
PubMed source DOI: 10.1091/mbc.3.1.29 PMC Full Text View journal details
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