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. 1995 Sep 2;130(6):1373–1385. doi: 10.1083/jcb.130.6.1373

Two microtubule-associated proteins required for anaphase spindle movement in Saccharomyces cerevisiae [published erratum appears in J Cell Biol 1995 Oct;131(2):561]

PMCID: PMC2120566  PMID: 7559759

Abstract

In many eucaryotic cells, the midzone of the mitotic spindle forms a distinct structure containing a specific set of proteins. We have isolated ASE1, a gene encoding a component of the Saccharomyces cerevisiae spindle midzone. Strains lacking both ASE1 and BIK1, which encodes an S. cerevisiae microtubule-associated protein, are inviable. The analysis of the phenotype of a bik1 ase1 conditional double mutant suggests that BIK1 and ASE1 are not required for the assembly of a bipolar spindle, but are essential for anaphase spindle elongation. The steady-state levels of Ase1p are regulated in a manner that is consistent with a function during anaphase: they are low in G1, accumulate to maximal levels after S phase and then drop as cells exit mitosis. Components of the spindle midzone may therefore be required in vivo for anaphase spindle movement. Additionally, anaphase spindle movement may depend on a dedicated set of genes whose expression is induced at G2/M.

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Selected References

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  1. Aist J. R., Berns M. W. Mechanics of chromosome separation during mitosis in Fusarium (Fungi imperfecti): new evidence from ultrastructural and laser microbeam experiments. J Cell Biol. 1981 Nov;91(2 Pt 1):446–458. doi: 10.1083/jcb.91.2.446. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alani E., Cao L., Kleckner N. A method for gene disruption that allows repeated use of URA3 selection in the construction of multiply disrupted yeast strains. Genetics. 1987 Aug;116(4):541–545. doi: 10.1534/genetics.112.541.test. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  4. Andreassen P. R., Palmer D. K., Wener M. H., Margolis R. L. Telophase disc: a new mammalian mitotic organelle that bisects telophase cells with a possible function in cytokinesis. J Cell Sci. 1991 Jul;99(Pt 3):523–534. doi: 10.1242/jcs.99.3.523. [DOI] [PubMed] [Google Scholar]
  5. Ault J. G., Rieder C. L. Centrosome and kinetochore movement during mitosis. Curr Opin Cell Biol. 1994 Feb;6(1):41–49. doi: 10.1016/0955-0674(94)90114-7. [DOI] [PubMed] [Google Scholar]
  6. Baum P., Yip C., Goetsch L., Byers B. A yeast gene essential for regulation of spindle pole duplication. Mol Cell Biol. 1988 Dec;8(12):5386–5397. doi: 10.1128/mcb.8.12.5386. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bender A., Pringle J. R. Use of a screen for synthetic lethal and multicopy suppressee mutants to identify two new genes involved in morphogenesis in Saccharomyces cerevisiae. Mol Cell Biol. 1991 Mar;11(3):1295–1305. doi: 10.1128/mcb.11.3.1295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Berlin V., Styles C. A., Fink G. R. 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):2573–2586. doi: 10.1083/jcb.111.6.2573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Bilbe G., Delabie J., Brüggen J., Richener H., Asselbergs F. A., Cerletti N., Sorg C., Odink K., Tarcsay L., Wiesendanger W. Restin: a novel intermediate filament-associated protein highly expressed in the Reed-Sternberg cells of Hodgkin's disease. EMBO J. 1992 Jun;11(6):2103–2113. doi: 10.1002/j.1460-2075.1992.tb05269.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Boeke J. D., Trueheart J., Natsoulis G., Fink G. R. 5-Fluoroorotic acid as a selective agent in yeast molecular genetics. Methods Enzymol. 1987;154:164–175. doi: 10.1016/0076-6879(87)54076-9. [DOI] [PubMed] [Google Scholar]
  11. Brown K. D., Coulson R. M., Yen T. J., Cleveland D. W. Cyclin-like accumulation and loss of the putative kinetochore motor CENP-E results from coupling continuous synthesis with specific degradation at the end of mitosis. J Cell Biol. 1994 Jun;125(6):1303–1312. doi: 10.1083/jcb.125.6.1303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Cande W. Z., McDonald K. L. In vitro reactivation of anaphase spindle elongation using isolated diatom spindles. Nature. 1985 Jul 11;316(6024):168–170. doi: 10.1038/316168a0. [DOI] [PubMed] [Google Scholar]
  14. Clark S. W., Meyer D. I. ACT3: a putative centractin homologue in S. cerevisiae is required for proper orientation of the mitotic spindle. J Cell Biol. 1994 Oct;127(1):129–138. doi: 10.1083/jcb.127.1.129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ding R., McDonald K. L., McIntosh J. R. Three-dimensional reconstruction and analysis of mitotic spindles from the yeast, Schizosaccharomyces pombe. J Cell Biol. 1993 Jan;120(1):141–151. doi: 10.1083/jcb.120.1.141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Earnshaw W. C., Mackay A. M. Role of nonhistone proteins in the chromosomal events of mitosis. FASEB J. 1994 Sep;8(12):947–956. doi: 10.1096/fasebj.8.12.8088460. [DOI] [PubMed] [Google Scholar]
  18. Elion E. A., Warner J. R. The major promoter element of rRNA transcription in yeast lies 2 kb upstream. Cell. 1984 Dec;39(3 Pt 2):663–673. doi: 10.1016/0092-8674(84)90473-2. [DOI] [PubMed] [Google Scholar]
  19. Eshel D., Urrestarazu L. A., Vissers S., Jauniaux J. C., van Vliet-Reedijk J. C., Planta R. J., Gibbons I. R. Cytoplasmic dynein is required for normal nuclear segregation in yeast. Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):11172–11176. doi: 10.1073/pnas.90.23.11172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Evan G. I., Lewis G. K., Ramsay G., Bishop J. M. Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product. Mol Cell Biol. 1985 Dec;5(12):3610–3616. doi: 10.1128/mcb.5.12.3610. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Garnier J., Osguthorpe D. J., Robson B. Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins. J Mol Biol. 1978 Mar 25;120(1):97–120. doi: 10.1016/0022-2836(78)90297-8. [DOI] [PubMed] [Google Scholar]
  22. Gelfand V. I., Bershadsky A. D. Microtubule dynamics: mechanism, regulation, and function. Annu Rev Cell Biol. 1991;7:93–116. doi: 10.1146/annurev.cb.07.110191.000521. [DOI] [PubMed] [Google Scholar]
  23. Gietz R. D., Sugino A. New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. Gene. 1988 Dec 30;74(2):527–534. doi: 10.1016/0378-1119(88)90185-0. [DOI] [PubMed] [Google Scholar]
  24. Guarente L. Synthetic enhancement in gene interaction: a genetic tool come of age. Trends Genet. 1993 Oct;9(10):362–366. doi: 10.1016/0168-9525(93)90042-g. [DOI] [PubMed] [Google Scholar]
  25. Hagan I., Yanagida M. Kinesin-related cut7 protein associates with mitotic and meiotic spindles in fission yeast. Nature. 1992 Mar 5;356(6364):74–76. doi: 10.1038/356074a0. [DOI] [PubMed] [Google Scholar]
  26. Hagan I., Yanagida M. Novel potential mitotic motor protein encoded by the fission yeast cut7+ gene. Nature. 1990 Oct 11;347(6293):563–566. doi: 10.1038/347563a0. [DOI] [PubMed] [Google Scholar]
  27. Hereford L. M., Osley M. A., Ludwig T. R., 2nd, McLaughlin C. S. Cell-cycle regulation of yeast histone mRNA. Cell. 1981 May;24(2):367–375. doi: 10.1016/0092-8674(81)90326-3. [DOI] [PubMed] [Google Scholar]
  28. Hoheisel J., Pohl F. M. Simplified preparation of unidirectional deletion clones. Nucleic Acids Res. 1986 Apr 25;14(8):3605–3605. doi: 10.1093/nar/14.8.3605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Hoyt M. A. Cellular roles of kinesin and related proteins. Curr Opin Cell Biol. 1994 Feb;6(1):63–68. doi: 10.1016/0955-0674(94)90117-1. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. 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]
  32. Hutter K. J., Eipel H. E. Microbial determinations by flow cytometry. J Gen Microbiol. 1979 Aug;113(2):369–375. doi: 10.1099/00221287-113-2-369. [DOI] [PubMed] [Google Scholar]
  33. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Koch C., Nasmyth K. Cell cycle regulated transcription in yeast. Curr Opin Cell Biol. 1994 Jun;6(3):451–459. doi: 10.1016/0955-0674(94)90039-6. [DOI] [PubMed] [Google Scholar]
  35. Koshland D., Kent J. C., Hartwell L. H. Genetic analysis of the mitotic transmission of minichromosomes. Cell. 1985 Feb;40(2):393–403. doi: 10.1016/0092-8674(85)90153-9. [DOI] [PubMed] [Google Scholar]
  36. Kunkel T. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci U S A. 1985 Jan;82(2):488–492. doi: 10.1073/pnas.82.2.488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Lawrence C. W. Classical mutagenesis techniques. Methods Enzymol. 1991;194:273–281. doi: 10.1016/0076-6879(91)94021-4. [DOI] [PubMed] [Google Scholar]
  38. Li Y. Y., Yeh E., Hays T., Bloom K. Disruption of mitotic spindle orientation in a yeast dynein mutant. Proc Natl Acad Sci U S A. 1993 Nov 1;90(21):10096–10100. doi: 10.1073/pnas.90.21.10096. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Lydall D., Ammerer G., Nasmyth K. A new role for MCM1 in yeast: cell cycle regulation of SW15 transcription. Genes Dev. 1991 Dec;5(12B):2405–2419. doi: 10.1101/gad.5.12b.2405. [DOI] [PubMed] [Google Scholar]
  40. Mastronarde D. N., McDonald K. L., Ding R., McIntosh J. R. Interpolar spindle microtubules in PTK cells. J Cell Biol. 1993 Dec;123(6 Pt 1):1475–1489. doi: 10.1083/jcb.123.6.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Masuda H., Hirano T., Yanagida M., Cande W. Z. In vitro reactivation of spindle elongation in fission yeast nuc2 mutant cells. J Cell Biol. 1990 Feb;110(2):417–425. doi: 10.1083/jcb.110.2.417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. May G. S., McGoldrick C. A., Holt C. L., Denison S. H. The bimB3 mutation of Aspergillus nidulans uncouples DNA replication from the completion of mitosis. J Biol Chem. 1992 Aug 5;267(22):15737–15743. [PubMed] [Google Scholar]
  43. McGrew J. T., Goetsch L., Byers B., Baum P. Requirement for ESP1 in the nuclear division of Saccharomyces cerevisiae. Mol Biol Cell. 1992 Dec;3(12):1443–1454. doi: 10.1091/mbc.3.12.1443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. McIntosh J. R., Hering G. E. Spindle fiber action and chromosome movement. Annu Rev Cell Biol. 1991;7:403–426. doi: 10.1146/annurev.cb.07.110191.002155. [DOI] [PubMed] [Google Scholar]
  45. McIntosh J. R., McDonald K. L., Edwards M. K., Ross B. M. Three-dimensional structure of the central mitotic spindle of Diatoma vulgare. J Cell Biol. 1979 Nov;83(2 Pt 1):428–442. doi: 10.1083/jcb.83.2.428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. McIntosh J. R., Roos U. P., Neighbors B., McDonald K. L. Architecture of the microtubule component of mitotic spindles from Dictyostelium discoideum. J Cell Sci. 1985 Apr;75:93–129. doi: 10.1242/jcs.75.1.93. [DOI] [PubMed] [Google Scholar]
  47. McMillan J. N., Tatchell K. The JNM1 gene in the yeast Saccharomyces cerevisiae is required for nuclear migration and spindle orientation during the mitotic cell cycle. J Cell Biol. 1994 Apr;125(1):143–158. doi: 10.1083/jcb.125.1.143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Mitchison T. J. Compare and contrast actin filaments and microtubules. Mol Biol Cell. 1992 Dec;3(12):1309–1315. doi: 10.1091/mbc.3.12.1309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Muhua L., Karpova T. S., Cooper J. A. A yeast actin-related protein homologous to that in vertebrate dynactin complex is important for spindle orientation and nuclear migration. Cell. 1994 Aug 26;78(4):669–679. doi: 10.1016/0092-8674(94)90531-2. [DOI] [PubMed] [Google Scholar]
  50. Nicklas R. B., Kubai D. F., Hays T. S. Spindle microtubules and their mechanical associations after micromanipulation in anaphase. J Cell Biol. 1982 Oct;95(1):91–104. doi: 10.1083/jcb.95.1.91. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Nislow C., Lombillo V. A., Kuriyama R., McIntosh J. R. A plus-end-directed motor enzyme that moves antiparallel microtubules in vitro localizes to the interzone of mitotic spindles. Nature. 1992 Oct 8;359(6395):543–547. doi: 10.1038/359543a0. [DOI] [PubMed] [Google Scholar]
  52. 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]
  53. Palmer R. E., Koval M., Koshland D. The dynamics of chromosome movement in the budding yeast Saccharomyces cerevisiae. J Cell Biol. 1989 Dec;109(6 Pt 2):3355–3366. doi: 10.1083/jcb.109.6.3355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Pasqualone D., Huffaker T. C. STU1, a suppressor of a beta-tubulin mutation, encodes a novel and essential component of the yeast mitotic spindle. J Cell Biol. 1994 Dec;127(6 Pt 2):1973–1984. doi: 10.1083/jcb.127.6.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Pierre P., Scheel J., Rickard J. E., Kreis T. E. CLIP-170 links endocytic vesicles to microtubules. Cell. 1992 Sep 18;70(6):887–900. doi: 10.1016/0092-8674(92)90240-d. [DOI] [PubMed] [Google Scholar]
  56. Pringle J. R., Adams A. E., Drubin D. G., Haarer B. K. Immunofluorescence methods for yeast. Methods Enzymol. 1991;194:565–602. doi: 10.1016/0076-6879(91)94043-c. [DOI] [PubMed] [Google Scholar]
  57. Riles L., Dutchik J. E., Baktha A., McCauley B. K., Thayer E. C., Leckie M. P., Braden V. V., Depke J. E., Olson M. V. Physical maps of the six smallest chromosomes of Saccharomyces cerevisiae at a resolution of 2.6 kilobase pairs. Genetics. 1993 May;134(1):81–150. doi: 10.1093/genetics/134.1.81. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. 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]
  59. 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]
  60. 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]
  61. Saunders W. S., Koshland D., Eshel D., Gibbons I. R., Hoyt M. A. Saccharomyces cerevisiae kinesin- and dynein-related proteins required for anaphase chromosome segregation. J Cell Biol. 1995 Feb;128(4):617–624. doi: 10.1083/jcb.128.4.617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  62. Schroer T. A. Structure, function and regulation of cytoplasmic dynein. Curr Opin Cell Biol. 1994 Feb;6(1):69–73. doi: 10.1016/0955-0674(94)90118-x. [DOI] [PubMed] [Google Scholar]
  63. 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]
  64. 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]
  65. Surana U., Amon A., Dowzer C., McGrew J., Byers B., Nasmyth K. Destruction of the CDC28/CLB mitotic kinase is not required for the metaphase to anaphase transition in budding yeast. EMBO J. 1993 May;12(5):1969–1978. doi: 10.1002/j.1460-2075.1993.tb05846.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  66. Trueheart J., Boeke J. D., Fink G. R. Two genes required for cell fusion during yeast conjugation: evidence for a pheromone-induced surface protein. Mol Cell Biol. 1987 Jul;7(7):2316–2328. doi: 10.1128/mcb.7.7.2316. [DOI] [PMC free article] [PubMed] [Google Scholar]
  67. Uzawa S., Samejima I., Hirano T., Tanaka K., Yanagida M. The fission yeast cut1+ gene regulates spindle pole body duplication and has homology to the budding yeast ESP1 gene. Cell. 1990 Sep 7;62(5):913–925. doi: 10.1016/0092-8674(90)90266-h. [DOI] [PubMed] [Google Scholar]
  68. Waters J. C., Cole R. W., Rieder C. L. The force-producing mechanism for centrosome separation during spindle formation in vertebrates is intrinsic to each aster. J Cell Biol. 1993 Jul;122(2):361–372. doi: 10.1083/jcb.122.2.361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  69. Winey M., Mamay C. L., O'Toole E. T., Mastronarde D. N., Giddings T. H., Jr, McDonald K. L., McIntosh J. R. Three-dimensional ultrastructural analysis of the Saccharomyces cerevisiae mitotic spindle. J Cell Biol. 1995 Jun;129(6):1601–1615. doi: 10.1083/jcb.129.6.1601. [DOI] [PMC free article] [PubMed] [Google Scholar]

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