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. 1996 Jul 2;134(2):455–464. doi: 10.1083/jcb.134.2.455

Anastral meiotic spindle morphogenesis: role of the non-claret disjunctional kinesin-like protein

PMCID: PMC2120873  PMID: 8707829

Abstract

We have used time-lapse laser scanning confocal microscopy to directly examine microtubule reorganization during meiotic spindle assembly in living Drosophila oocytes. These studies indicate that the bipolarity of the meiosis I spindle is not the result of a duplication and separation of centrosomal microtubule organizing centers (MTOCs). Instead, microtubules first associate with a tight chromatin mass, and then bundle to form a bipolar spindle that lacks asters. Analysis of mutant oocytes indicates that the Non-Claret Disjunctional (NCD) kinesin-like protein is required for normal spindle assembly kinetics and stabilization of the spindle during metaphase arrest. Immunolocalization analyses demonstrate that NCD is associated with spindle microtubules, and that the centrosomal components gamma- tubulin, CP-190, and CP-60 are not concentrated at the meiotic spindle poles. Based on these observations, we propose that microtubule bundling by the NCD kinesin-like protein promotes assembly of a stable bipolar spindle in the absence of typical MTOCs.

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Selected References

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  1. Chandra R., Salmon E. D., Erickson H. P., Lockhart A., Endow S. A. Structural and functional domains of the Drosophila ncd microtubule motor protein. J Biol Chem. 1993 Apr 25;268(12):9005–9013. [PubMed] [Google Scholar]
  2. Davis D. G. Chromosome Behavior under the Influence of Claret-Nondisjunctional in DROSOPHILA MELANOGASTER. Genetics. 1969 Mar;61(3):577–594. doi: 10.1093/genetics/61.3.577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Endow S. A., Chandra R., Komma D. J., Yamamoto A. H., Salmon E. D. Mutants of the Drosophila ncd microtubule motor protein cause centrosomal and spindle pole defects in mitosis. J Cell Sci. 1994 Apr;107(Pt 4):859–867. doi: 10.1242/jcs.107.4.859. [DOI] [PubMed] [Google Scholar]
  4. Endow S. A., Henikoff S., Soler-Niedziela L. Mediation of meiotic and early mitotic chromosome segregation in Drosophila by a protein related to kinesin. Nature. 1990 May 3;345(6270):81–83. doi: 10.1038/345081a0. [DOI] [PubMed] [Google Scholar]
  5. Hatsumi M., Endow S. A. Mutants of the microtubule motor protein, nonclaret disjunctional, affect spindle structure and chromosome movement in meiosis and mitosis. J Cell Sci. 1992 Mar;101(Pt 3):547–559. doi: 10.1242/jcs.101.3.547. [DOI] [PubMed] [Google Scholar]
  6. Hatsumi M., Endow S. A. The Drosophila ncd microtubule motor protein is spindle-associated in meiotic and mitotic cells. J Cell Sci. 1992 Dec;103(Pt 4):1013–1020. doi: 10.1242/jcs.103.4.1013. [DOI] [PubMed] [Google Scholar]
  7. Hawley R. S. Genetic and molecular analysis of a simple disjunctional system in Drosophila melanogaster. Prog Clin Biol Res. 1989;311:277–302. [PubMed] [Google Scholar]
  8. Hawley R. S., McKim K. S., Arbel T. Meiotic segregation in Drosophila melanogaster females: molecules, mechanisms, and myths. Annu Rev Genet. 1993;27:281–317. doi: 10.1146/annurev.ge.27.120193.001433. [DOI] [PubMed] [Google Scholar]
  9. Kellogg D. R., Alberts B. M. Purification of a multiprotein complex containing centrosomal proteins from the Drosophila embryo by chromatography with low-affinity polyclonal antibodies. Mol Biol Cell. 1992 Jan;3(1):1–11. doi: 10.1091/mbc.3.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kellogg D. R., Field C. M., Alberts B. M. 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):2977–2991. doi: 10.1083/jcb.109.6.2977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kellogg D. R., Mitchison T. J., Alberts B. M. Behaviour of microtubules and actin filaments in living Drosophila embryos. Development. 1988 Aug;103(4):675–686. doi: 10.1242/dev.103.4.675. [DOI] [PubMed] [Google Scholar]
  12. Kimble M., Church K. Meiosis and early cleavage in Drosophila melanogaster eggs: effects of the claret-non-disjunctional mutation. J Cell Sci. 1983 Jul;62:301–318. doi: 10.1242/jcs.62.1.301. [DOI] [PubMed] [Google Scholar]
  13. Kuriyama R., Kofron M., Essner R., Kato T., Dragas-Granoic S., Omoto C. K., Khodjakov A. Characterization of a minus end-directed kinesin-like motor protein from cultured mammalian cells. J Cell Biol. 1995 May;129(4):1049–1059. doi: 10.1083/jcb.129.4.1049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. McDonald H. B., Goldstein L. S. Identification and characterization of a gene encoding a kinesin-like protein in Drosophila. Cell. 1990 Jun 15;61(6):991–1000. doi: 10.1016/0092-8674(90)90064-l. [DOI] [PubMed] [Google Scholar]
  15. McDonald H. B., Stewart R. J., Goldstein L. S. The kinesin-like ncd protein of Drosophila is a minus end-directed microtubule motor. Cell. 1990 Dec 21;63(6):1159–1165. doi: 10.1016/0092-8674(90)90412-8. [DOI] [PubMed] [Google Scholar]
  16. McKim K. S., Hawley R. S. Chromosomal control of meiotic cell division. Science. 1995 Dec 8;270(5242):1595–1601. doi: 10.1126/science.270.5242.1595. [DOI] [PubMed] [Google Scholar]
  17. McKim K. S., Jang J. K., Theurkauf W. E., Hawley R. S. Mechanical basis of meiotic metaphase arrest. Nature. 1993 Mar 25;362(6418):364–366. doi: 10.1038/362364a0. [DOI] [PubMed] [Google Scholar]
  18. Miyazaki W. Y., Orr-Weaver T. L. Sister-chromatid cohesion in mitosis and meiosis. Annu Rev Genet. 1994;28:167–187. doi: 10.1146/annurev.ge.28.120194.001123. [DOI] [PubMed] [Google Scholar]
  19. NOVITSKI E. AN ALTERNATIVE TO THE DISTRIBUTIVE PAIRING HYPOTHESIS IN DROSOPHILA. Genetics. 1964 Dec;50:1449–1451. doi: 10.1093/genetics/50.6.1449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nicklas R. B. Chromosome micromanipulation. II. Induced reorientation and the experimental control of segregation in meiosis. Chromosoma. 1967;21(1):17–50. doi: 10.1007/BF00330545. [DOI] [PubMed] [Google Scholar]
  21. Nicklas R. B. Chromosome segregation mechanisms. Genetics. 1974 Sep;78(1):205–213. doi: 10.1093/genetics/78.1.205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Nicklas R. B., Koch C. A. Chromosome micromanipulation. 3. Spindle fiber tension and the reorientation of mal-oriented chromosomes. J Cell Biol. 1969 Oct;43(1):40–50. doi: 10.1083/jcb.43.1.40. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nicklas R. B., Staehly C. A. Chromosome micromanipulation. I. The mechanics of chromosome attachment to the spindle. Chromosoma. 1967;21(1):1–16. doi: 10.1007/BF00330544. [DOI] [PubMed] [Google Scholar]
  24. Nicklas R. B. The motor for poleward chromosome movement in anaphase is in or near the kinetochore. J Cell Biol. 1989 Nov;109(5):2245–2255. doi: 10.1083/jcb.109.5.2245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. 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]
  27. Raff J. W., Kellogg D. R., Alberts B. M. Drosophila gamma-tubulin is part of a complex containing two previously identified centrosomal MAPs. J Cell Biol. 1993 May;121(4):823–835. doi: 10.1083/jcb.121.4.823. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rieder C. L., Salmon E. D. Motile kinetochores and polar ejection forces dictate chromosome position on the vertebrate mitotic spindle. J Cell Biol. 1994 Feb;124(3):223–233. doi: 10.1083/jcb.124.3.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Stewart R. J., Thaler J. P., Goldstein L. S. Direction of microtubule movement is an intrinsic property of the motor domains of kinesin heavy chain and Drosophila ncd protein. Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):5209–5213. doi: 10.1073/pnas.90.11.5209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Theurkauf W. E., Hawley R. S. Meiotic spindle assembly in Drosophila females: behavior of nonexchange chromosomes and the effects of mutations in the nod kinesin-like protein. J Cell Biol. 1992 Mar;116(5):1167–1180. doi: 10.1083/jcb.116.5.1167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Theurkauf W. E. Immunofluorescence analysis of the cytoskeleton during oogenesis and early embryogenesis. Methods Cell Biol. 1994;44:489–505. doi: 10.1016/s0091-679x(08)60928-0. [DOI] [PubMed] [Google Scholar]
  32. Theurkauf W. E. Premature microtubule-dependent cytoplasmic streaming in cappuccino and spire mutant oocytes. Science. 1994 Sep 30;265(5181):2093–2096. doi: 10.1126/science.8091233. [DOI] [PubMed] [Google Scholar]
  33. Vernos I., Karsenti E. Chromosomes take the lead in spindle assembly. Trends Cell Biol. 1995 Aug;5(8):297–301. doi: 10.1016/s0962-8924(00)89045-5. [DOI] [PubMed] [Google Scholar]
  34. Vernos I., Raats J., Hirano T., Heasman J., Karsenti E., Wylie C. Xklp1, a chromosomal Xenopus kinesin-like protein essential for spindle organization and chromosome positioning. Cell. 1995 Apr 7;81(1):117–127. doi: 10.1016/0092-8674(95)90376-3. [DOI] [PubMed] [Google Scholar]
  35. Wald H. Cytologic Studies on the Abnormal Development of the Eggs of the Claret Mutant Type of Drosophila Simulans. Genetics. 1936 May;21(3):264–281. doi: 10.1093/genetics/21.3.264. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Walker R. A., Salmon E. D., Endow S. A. The Drosophila claret segregation protein is a minus-end directed motor molecule. Nature. 1990 Oct 25;347(6295):780–782. doi: 10.1038/347780a0. [DOI] [PubMed] [Google Scholar]
  37. Yamamoto A. H., Komma D. J., Shaffer C. D., Pirrotta V., Endow S. A. The claret locus in Drosophila encodes products required for eyecolor and for meiotic chromosome segregation. EMBO J. 1989 Dec 1;8(12):3543–3552. doi: 10.1002/j.1460-2075.1989.tb08526.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Zheng Y., Wong M. L., Alberts B., Mitchison T. Nucleation of microtubule assembly by a gamma-tubulin-containing ring complex. Nature. 1995 Dec 7;378(6557):578–583. doi: 10.1038/378578a0. [DOI] [PubMed] [Google Scholar]

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