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. 1995 Dec 2;131(6):1775–1788. doi: 10.1083/jcb.131.6.1775

A highly divergent gamma-tubulin gene is essential for cell growth and proper microtubule organization in Saccharomyces cerevisiae

PMCID: PMC2120663  PMID: 8557744

Abstract

A Saccharomyces cerevisiae gamma-tubulin-related gene, TUB4, has been characterized. The predicted amino acid sequence of the Tub4 protein (Tub4p) is 29-38% identical to members of the gamma-tubulin family. Indirect immunofluorescence experiments using a strain containing an epitope-tagged Tub4p indicate that Tub4p resides at the spindle pole body throughout the yeast cell cycle. Deletion of the TUB4 gene indicates that Tub4p is essential for yeast cell growth. Tub4p-depleted cells arrest during nuclear division; most arrested cells contain a large bud, replicated DNA, and a single nucleus. Immunofluorescence and nuclear staining experiments indicate that cells depleted of Tub4p contain defects in the organization of both cytoplasmic and nuclear microtubule arrays; such cells exhibit nuclear migration failure, defects in spindle formation, and/or aberrantly long cytoplasmic microtubule arrays. These data indicate that the S. cerevisiae gamma- tubulin protein is an important SPB component that organizes both cytoplasmic and nuclear microtubule arrays.

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Selected References

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  1. Adams A. E., Pringle J. R. Relationship of actin and tubulin distribution to bud growth in wild-type and morphogenetic-mutant Saccharomyces cerevisiae. J Cell Biol. 1984 Mar;98(3):934–945. doi: 10.1083/jcb.98.3.934. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baudin A., Ozier-Kalogeropoulos O., Denouel A., Lacroute F., Cullin C. A simple and efficient method for direct gene deletion in Saccharomyces cerevisiae. Nucleic Acids Res. 1993 Jul 11;21(14):3329–3330. doi: 10.1093/nar/21.14.3329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baum P., Furlong C., Byers B. Yeast gene required for spindle pole body duplication: homology of its product with Ca2+-binding proteins. Proc Natl Acad Sci U S A. 1986 Aug;83(15):5512–5516. doi: 10.1073/pnas.83.15.5512. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bond J. F., Fridovich-Keil J. L., Pillus L., Mulligan R. C., Solomon F. A chicken-yeast chimeric beta-tubulin protein is incorporated into mouse microtubules in vivo. Cell. 1986 Feb 14;44(3):461–468. doi: 10.1016/0092-8674(86)90467-8. [DOI] [PubMed] [Google Scholar]
  5. Brinkley B. R. Microtubule organizing centers. Annu Rev Cell Biol. 1985;1:145–172. doi: 10.1146/annurev.cb.01.110185.001045. [DOI] [PubMed] [Google Scholar]
  6. Byers B., Goetsch L. Behavior of spindles and spindle plaques in the cell cycle and conjugation of Saccharomyces cerevisiae. J Bacteriol. 1975 Oct;124(1):511–523. doi: 10.1128/jb.124.1.511-523.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Byers B., Goetsch L. Duplication of spindle plaques and integration of the yeast cell cycle. Cold Spring Harb Symp Quant Biol. 1974;38:123–131. doi: 10.1101/sqb.1974.038.01.016. [DOI] [PubMed] [Google Scholar]
  8. Chen D. C., Yang B. C., Kuo T. T. One-step transformation of yeast in stationary phase. Curr Genet. 1992 Jan;21(1):83–84. doi: 10.1007/BF00318659. [DOI] [PubMed] [Google Scholar]
  9. Conde J., Fink G. R. A mutant of Saccharomyces cerevisiae defective for nuclear fusion. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3651–3655. doi: 10.1073/pnas.73.10.3651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fuchs U., Moepps B., Maucher H. P., Schraudolf H. 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. doi: 10.1007/BF00019306. [DOI] [PubMed] [Google Scholar]
  11. Gard D. L. Gamma-tubulin is asymmetrically distributed in the cortex of Xenopus oocytes. Dev Biol. 1994 Jan;161(1):131–140. doi: 10.1006/dbio.1994.1015. [DOI] [PubMed] [Google Scholar]
  12. Geiser J. R., Sundberg H. A., Chang B. H., Muller E. G., Davis T. N. The essential mitotic target of calmodulin is the 110-kilodalton component of the spindle pole body in Saccharomyces cerevisiae. Mol Cell Biol. 1993 Dec;13(12):7913–7924. doi: 10.1128/mcb.13.12.7913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hartwell L. H., Mortimer R. K., Culotti J., Culotti M. Genetic Control of the Cell Division Cycle in Yeast: V. Genetic Analysis of cdc Mutants. Genetics. 1973 Jun;74(2):267–286. doi: 10.1093/genetics/74.2.267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Horio T., Uzawa S., Jung M. K., Oakley B. R., Tanaka K., Yanagida M. 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. doi: 10.1242/jcs.99.4.693. [DOI] [PubMed] [Google Scholar]
  15. Hoyt M. A., He L., Loo K. K., Saunders W. S. Two Saccharomyces cerevisiae kinesin-related gene products required for mitotic spindle assembly. J Cell Biol. 1992 Jul;118(1):109–120. doi: 10.1083/jcb.118.1.109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hoyt M. A., Stearns T., Botstein D. Chromosome instability mutants of Saccharomyces cerevisiae that are defective in microtubule-mediated processes. Mol Cell Biol. 1990 Jan;10(1):223–234. doi: 10.1128/mcb.10.1.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Huffaker T. C., Thomas J. H., Botstein D. Diverse effects of beta-tubulin mutations on microtubule formation and function. J Cell Biol. 1988 Jun;106(6):1997–2010. doi: 10.1083/jcb.106.6.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hutter K. J., Eipel H. E. Flow cytometric determinations of cellular substances in algae, bacteria, moulds and yeasts. Antonie Van Leeuwenhoek. 1978;44(3-4):269–282. doi: 10.1007/BF00394305. [DOI] [PubMed] [Google Scholar]
  19. Jacobs C. W., Adams A. E., Szaniszlo P. J., Pringle J. R. Functions of microtubules in the Saccharomyces cerevisiae cell cycle. J Cell Biol. 1988 Oct;107(4):1409–1426. doi: 10.1083/jcb.107.4.1409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Johnston M., Davis R. W. Sequences that regulate the divergent GAL1-GAL10 promoter in Saccharomyces cerevisiae. Mol Cell Biol. 1984 Aug;4(8):1440–1448. doi: 10.1128/mcb.4.8.1440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Joshi H. C. Gamma-tubulin: the hub of cellular microtubule assemblies. Bioessays. 1993 Oct;15(10):637–643. doi: 10.1002/bies.950151002. [DOI] [PubMed] [Google Scholar]
  22. Joshi H. C., Palacios M. J., McNamara L., Cleveland D. W. Gamma-tubulin is a centrosomal protein required for cell cycle-dependent microtubule nucleation. Nature. 1992 Mar 5;356(6364):80–83. doi: 10.1038/356080a0. [DOI] [PubMed] [Google Scholar]
  23. Julian M., Tollon Y., Lajoie-Mazenc I., Moisand A., Mazarguil H., Puget A., Wright M. gamma-Tubulin participates in the formation of the midbody during cytokinesis in mammalian cells. J Cell Sci. 1993 May;105(Pt 1):145–156. doi: 10.1242/jcs.105.1.145. [DOI] [PubMed] [Google Scholar]
  24. Kalt A., Schliwa M. Molecular components of the centrosome. Trends Cell Biol. 1993 Apr;3(4):118–128. doi: 10.1016/0962-8924(93)90174-y. [DOI] [PubMed] [Google Scholar]
  25. Kilmartin J. V., Adams A. E. Structural rearrangements of tubulin and actin during the cell cycle of the yeast Saccharomyces. J Cell Biol. 1984 Mar;98(3):922–933. doi: 10.1083/jcb.98.3.922. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kilmartin J. V., Dyos S. L., Kershaw D., Finch J. T. A spacer protein in the Saccharomyces cerevisiae spindle poly body whose transcript is cell cycle-regulated. J Cell Biol. 1993 Dec;123(5):1175–1184. doi: 10.1083/jcb.123.5.1175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kilmartin J. V., Wright B., Milstein C. Rat monoclonal antitubulin antibodies derived by using a new nonsecreting rat cell line. J Cell Biol. 1982 Jun;93(3):576–582. doi: 10.1083/jcb.93.3.576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Lajoie-Mazenc I., Tollon Y., Detraves C., Julian M., Moisand A., Gueth-Hallonet C., Debec A., Salles-Passador I., Puget A., Mazarguil H. Recruitment of antigenic gamma-tubulin during mitosis in animal cells: presence of gamma-tubulin in the mitotic spindle. J Cell Sci. 1994 Oct;107(Pt 10):2825–2837. doi: 10.1242/jcs.107.10.2825. [DOI] [PubMed] [Google Scholar]
  29. Liang A., Heckmann K. The macronuclear gamma-tubulin-encoding gene of Euplotes octocarinatus contains two introns and an in-frame TGA. Gene. 1993 Dec 22;136(1-2):319–322. doi: 10.1016/0378-1119(93)90487-n. [DOI] [PubMed] [Google Scholar]
  30. Lillie S. H., Brown S. S. Suppression of a myosin defect by a kinesin-related gene. Nature. 1992 Mar 26;356(6367):358–361. doi: 10.1038/356358a0. [DOI] [PubMed] [Google Scholar]
  31. Liu B., Joshi H. C., Wilson T. J., Silflow C. D., Palevitz B. A., Snustad D. P. gamma-Tubulin in Arabidopsis: gene sequence, immunoblot, and immunofluorescence studies. Plant Cell. 1994 Feb;6(2):303–314. doi: 10.1105/tpc.6.2.303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Lowndes N. F., Johnson A. L., Johnston L. H. Coordination of expression of DNA synthesis genes in budding yeast by a cell-cycle regulated trans factor. Nature. 1991 Mar 21;350(6315):247–250. doi: 10.1038/350247a0. [DOI] [PubMed] [Google Scholar]
  33. Lowndes N. F., Johnston L. H. Parallel pathways of cell cycle-regulated gene expression. Trends Genet. 1992 Mar;8(3):79–81. doi: 10.1016/0168-9525(92)90183-5. [DOI] [PubMed] [Google Scholar]
  34. Luo H., Perlin M. H. The gamma-tubulin-encoding gene from the basidiomycete fungus, Ustilago violacea, has a long 5'-untranslated region. Gene. 1993 Dec 31;137(2):187–194. doi: 10.1016/0378-1119(93)90005-n. [DOI] [PubMed] [Google Scholar]
  35. Maessen S., Wesseling J. G., Smits M. A., Konings R. N., Schoenmakers J. G. The gamma-tubulin gene of the malaria parasite Plasmodium falciparum. Mol Biochem Parasitol. 1993 Jul;60(1):27–35. doi: 10.1016/0166-6851(93)90025-s. [DOI] [PubMed] [Google Scholar]
  36. Masuda H., Sevik M., Cande W. Z. 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):1055–1066. doi: 10.1083/jcb.117.5.1055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Neff N. F., Thomas J. H., Grisafi P., Botstein D. Isolation of the beta-tubulin gene from yeast and demonstration of its essential function in vivo. Cell. 1983 May;33(1):211–219. doi: 10.1016/0092-8674(83)90350-1. [DOI] [PubMed] [Google Scholar]
  38. Oakley B. R. Gamma-tubulin: the microtubule organizer? Trends Cell Biol. 1992 Jan;2(1):1–5. doi: 10.1016/0962-8924(92)90125-7. [DOI] [PubMed] [Google Scholar]
  39. Oakley B. R., Oakley C. E., Yoon Y., Jung M. K. 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):1289–1301. doi: 10.1016/0092-8674(90)90693-9. [DOI] [PubMed] [Google Scholar]
  40. Oakley C. E., Oakley B. R. Identification of gamma-tubulin, a new member of the tubulin superfamily encoded by mipA gene of Aspergillus nidulans. Nature. 1989 Apr 20;338(6217):662–664. doi: 10.1038/338662a0. [DOI] [PubMed] [Google Scholar]
  41. Osborne M. A., Schlenstedt G., Jinks T., Silver P. A. Nuf2, a spindle pole body-associated protein required for nuclear division in yeast. J Cell Biol. 1994 May;125(4):853–866. doi: 10.1083/jcb.125.4.853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Page B. D., Snyder M. CIK1: a developmentally regulated spindle pole body-associated protein important for microtubule functions in Saccharomyces cerevisiae. Genes Dev. 1992 Aug;6(8):1414–1429. doi: 10.1101/gad.6.8.1414. [DOI] [PubMed] [Google Scholar]
  43. Page B. D., Snyder M. Chromosome segregation in yeast. Annu Rev Microbiol. 1993;47:231–261. doi: 10.1146/annurev.mi.47.100193.001311. [DOI] [PubMed] [Google Scholar]
  44. Palmer R. E., Sullivan D. S., Huffaker T., Koshland D. Role of astral microtubules and actin in spindle orientation and migration in the budding yeast, Saccharomyces cerevisiae. J Cell Biol. 1992 Nov;119(3):583–593. doi: 10.1083/jcb.119.3.583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Roof D. M., Meluh P. B., Rose M. D. Kinesin-related proteins required for assembly of the mitotic spindle. J Cell Biol. 1992 Jul;118(1):95–108. doi: 10.1083/jcb.118.1.95. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Rose M. D., Biggins S., Satterwhite L. L. Unravelling the tangled web at the microtubule-organizing center. Curr Opin Cell Biol. 1993 Feb;5(1):105–115. doi: 10.1016/s0955-0674(05)80015-8. [DOI] [PubMed] [Google Scholar]
  47. Rose M. D., Fink G. R. KAR1, a gene required for function of both intranuclear and extranuclear microtubules in yeast. Cell. 1987 Mar 27;48(6):1047–1060. doi: 10.1016/0092-8674(87)90712-4. [DOI] [PubMed] [Google Scholar]
  48. Rout M. P., Kilmartin J. V. Components of the yeast spindle and spindle pole body. J Cell Biol. 1990 Nov;111(5 Pt 1):1913–1927. doi: 10.1083/jcb.111.5.1913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Rout M. P., Kilmartin J. V. Yeast spindle pole body components. Cold Spring Harb Symp Quant Biol. 1991;56:687–692. doi: 10.1101/sqb.1991.056.01.077. [DOI] [PubMed] [Google Scholar]
  50. Saunders W. S., Hoyt M. A. Kinesin-related proteins required for structural integrity of the mitotic spindle. Cell. 1992 Aug 7;70(3):451–458. doi: 10.1016/0092-8674(92)90169-d. [DOI] [PubMed] [Google Scholar]
  51. Schatz P. J., Pillus L., Grisafi P., Solomon F., Botstein D. Two functional alpha-tubulin genes of the yeast Saccharomyces cerevisiae encode divergent proteins. Mol Cell Biol. 1986 Nov;6(11):3711–3721. doi: 10.1128/mcb.6.11.3711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Schatz P. J., Solomon F., Botstein D. Genetically essential and nonessential alpha-tubulin genes specify functionally interchangeable proteins. Mol Cell Biol. 1986 Nov;6(11):3722–3733. doi: 10.1128/mcb.6.11.3722. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Schneider B. L., Seufert W., Steiner B., Yang Q. H., Futcher A. B. Use of polymerase chain reaction epitope tagging for protein tagging in Saccharomyces cerevisiae. Yeast. 1995 Oct;11(13):1265–1274. doi: 10.1002/yea.320111306. [DOI] [PubMed] [Google Scholar]
  54. Sethi N., Monteagudo M. C., Koshland D., Hogan E., Burke D. J. The CDC20 gene product of Saccharomyces cerevisiae, a beta-transducin homolog, is required for a subset of microtubule-dependent cellular processes. Mol Cell Biol. 1991 Nov;11(11):5592–5602. doi: 10.1128/mcb.11.11.5592. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Sikorski R. S., Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. doi: 10.1093/genetics/122.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Snyder M., Davis R. W. SPA1: a gene important for chromosome segregation and other mitotic functions in S. cerevisiae. Cell. 1988 Sep 9;54(6):743–754. doi: 10.1016/s0092-8674(88)90977-4. [DOI] [PubMed] [Google Scholar]
  57. Snyder M., Gehrung S., Page B. D. Studies concerning the temporal and genetic control of cell polarity in Saccharomyces cerevisiae. J Cell Biol. 1991 Aug;114(3):515–532. doi: 10.1083/jcb.114.3.515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Snyder M. The spindle pole body of yeast. Chromosoma. 1994 Oct;103(6):369–380. doi: 10.1007/BF00362281. [DOI] [PubMed] [Google Scholar]
  59. Spang A., Courtney I., Fackler U., Matzner M., Schiebel E. The calcium-binding protein cell division cycle 31 of Saccharomyces cerevisiae is a component of the half bridge of the spindle pole body. J Cell Biol. 1993 Oct;123(2):405–416. doi: 10.1083/jcb.123.2.405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Spang A., Courtney I., Grein K., Matzner M., Schiebel E. The Cdc31p-binding protein Kar1p is a component of the half bridge of the yeast spindle pole body. J Cell Biol. 1995 Mar;128(5):863–877. doi: 10.1083/jcb.128.5.863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Spencer F., Gerring S. L., Connelly C., Hieter P. Mitotic chromosome transmission fidelity mutants in Saccharomyces cerevisiae. Genetics. 1990 Feb;124(2):237–249. doi: 10.1093/genetics/124.2.237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  62. Stearns T., Evans L., Kirschner M. Gamma-tubulin is a highly conserved component of the centrosome. Cell. 1991 May 31;65(5):825–836. doi: 10.1016/0092-8674(91)90390-k. [DOI] [PubMed] [Google Scholar]
  63. Stearns T., Hoyt M. A., Botstein D. Yeast mutants sensitive to antimicrotubule drugs define three genes that affect microtubule function. Genetics. 1990 Feb;124(2):251–262. doi: 10.1093/genetics/124.2.251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  64. Stirling D. A., Welch K. A., Stark M. J. Interaction with calmodulin is required for the function of Spc110p, an essential component of the yeast spindle pole body. EMBO J. 1994 Sep 15;13(18):4329–4342. doi: 10.1002/j.1460-2075.1994.tb06753.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  65. Sullivan D. S., Huffaker T. C. Astral microtubules are not required for anaphase B in Saccharomyces cerevisiae. J Cell Biol. 1992 Oct;119(2):379–388. doi: 10.1083/jcb.119.2.379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  66. Sulston J., Du Z., Thomas K., Wilson R., Hillier L., Staden R., Halloran N., Green P., Thierry-Mieg J., Qiu L. The C. elegans genome sequencing project: a beginning. Nature. 1992 Mar 5;356(6364):37–41. doi: 10.1038/356037a0. [DOI] [PubMed] [Google Scholar]
  67. Sun G. H., Hirata A., Ohya Y., Anraku Y. Mutations in yeast calmodulin cause defects in spindle pole body functions and nuclear integrity. J Cell Biol. 1992 Dec;119(6):1625–1639. doi: 10.1083/jcb.119.6.1625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  68. Sunkel C. E., Gomes R., Sampaio P., Perdigão J., González C. 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. doi: 10.1002/j.1460-2075.1995.tb06972.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  69. Vallen E. A., Scherson T. Y., Roberts T., van Zee K., Rose M. D. Asymmetric mitotic segregation of the yeast spindle pole body. Cell. 1992 May 1;69(3):505–515. doi: 10.1016/0092-8674(92)90451-h. [DOI] [PubMed] [Google Scholar]
  70. Wilson I. A., Niman H. L., Houghten R. A., Cherenson A. R., Connolly M. L., Lerner R. A. The structure of an antigenic determinant in a protein. Cell. 1984 Jul;37(3):767–778. doi: 10.1016/0092-8674(84)90412-4. [DOI] [PubMed] [Google Scholar]
  71. Winey M., Byers B. Assembly and functions of the spindle pole body in budding yeast. Trends Genet. 1993 Sep;9(9):300–304. doi: 10.1016/0168-9525(93)90247-f. [DOI] [PubMed] [Google Scholar]
  72. Winey M., Goetsch L., Baum P., Byers B. MPS1 and MPS2: novel yeast genes defining distinct steps of spindle pole body duplication. J Cell Biol. 1991 Aug;114(4):745–754. doi: 10.1083/jcb.114.4.745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  73. Zheng Y., Jung M. K., Oakley B. R. Gamma-tubulin is present in Drosophila melanogaster and Homo sapiens and is associated with the centrosome. Cell. 1991 May 31;65(5):817–823. doi: 10.1016/0092-8674(91)90389-g. [DOI] [PubMed] [Google Scholar]

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