Abstract
The female meiotic spindle lacks a centrosome or microtubule-organizing center in many organisms. During cell division, these spindles are organized by the chromosomes and microtubule-associated proteins. Previous studies in Drosophila melanogaster implicated at least one kinesin motor protein, NCD, in tapering the microtubules into a bipolar spindle. We have identified a second Drosophila kinesin-like protein, SUB, that is required for meiotic spindle function. At meiosis I in males and females, sub mutations affect only the segregation of homologous chromosomes. In female meiosis, sub mutations have a similar phenotype to ncd; even though chromosomes are joined by chiasmata they fail to segregate at meiosis I. Cytological analyses have revealed that sub is required for bipolar spindle formation. In sub mutations, we observed spindles that were unipolar, multipolar, or frayed with no defined poles. On the basis of these phenotypes and the observation that sub mutations genetically interact with ncd, we propose that SUB is one member of a group of microtubule-associated proteins required for bipolar spindle assembly in the absence of the centrosomes. sub is also required for the early embryonic divisions but is otherwise dispensable for most mitotic divisions.
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- Adams R. R., Tavares A. A., Salzberg A., Bellen H. J., Glover D. M. pavarotti encodes a kinesin-like protein required to organize the central spindle and contractile ring for cytokinesis. Genes Dev. 1998 May 15;12(10):1483–1494. doi: 10.1101/gad.12.10.1483. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Boleti H., Karsenti E., Vernos I. Xklp2, a novel Xenopus centrosomal kinesin-like protein required for centrosome separation during mitosis. Cell. 1996 Jan 12;84(1):49–59. doi: 10.1016/s0092-8674(00)80992-7. [DOI] [PubMed] [Google Scholar]
- Brent A. E., MacQueen A., Hazelrigg T. The Drosophila wispy gene is required for RNA localization and other microtubule-based events of meiosis and early embryogenesis. Genetics. 2000 Apr;154(4):1649–1662. doi: 10.1093/genetics/154.4.1649. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Case R. B., Rice S., Hart C. L., Ly B., Vale R. D. Role of the kinesin neck linker and catalytic core in microtubule-based motility. Curr Biol. 2000 Feb 10;10(3):157–160. doi: 10.1016/s0960-9822(00)00316-x. [DOI] [PubMed] [Google Scholar]
- Cross R. A. Molecular motors: Kinesin's string variable. Curr Biol. 2001 Feb 20;11(4):R147–R149. doi: 10.1016/s0960-9822(01)00064-1. [DOI] [PubMed] [Google Scholar]
- Cullen C. F., Ohkura H. Msps protein is localized to acentrosomal poles to ensure bipolarity of Drosophila meiotic spindles. Nat Cell Biol. 2001 Jul;3(7):637–642. doi: 10.1038/35083025. [DOI] [PubMed] [Google Scholar]
- Dernburg A. F., Sedat J. W., Hawley R. S. Direct evidence of a role for heterochromatin in meiotic chromosome segregation. Cell. 1996 Jul 12;86(1):135–146. doi: 10.1016/s0092-8674(00)80084-7. [DOI] [PubMed] [Google Scholar]
- Echard A., Jollivet F., Martinez O., Lacapère J. J., Rousselet A., Janoueix-Lerosey I., Goud B. Interaction of a Golgi-associated kinesin-like protein with Rab6. Science. 1998 Jan 23;279(5350):580–585. doi: 10.1126/science.279.5350.580. [DOI] [PubMed] [Google Scholar]
- Fontijn R. D., Goud B., Echard A., Jollivet F., van Marle J., Pannekoek H., Horrevoets A. J. The human kinesin-like protein RB6K is under tight cell cycle control and is essential for cytokinesis. Mol Cell Biol. 2001 Apr;21(8):2944–2955. doi: 10.1128/MCB.21.8.2944-2955.2001. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Hawley R. S., Theurkauf W. E. Requiem for distributive segregation: achiasmate segregation in Drosophila females. Trends Genet. 1993 Sep;9(9):310–317. doi: 10.1016/0168-9525(93)90249-h. [DOI] [PubMed] [Google Scholar]
- Heald R., Tournebize R., Blank T., Sandaltzopoulos R., Becker P., Hyman A., Karsenti E. Self-organization of microtubules into bipolar spindles around artificial chromosomes in Xenopus egg extracts. Nature. 1996 Aug 1;382(6590):420–425. doi: 10.1038/382420a0. [DOI] [PubMed] [Google Scholar]
- Kim A. J., Endow S. A. A kinesin family tree. J Cell Sci. 2000 Nov;113(Pt 21):3681–3682. doi: 10.1242/jcs.113.21.3681. [DOI] [PubMed] [Google Scholar]
- Knowles B. A., Hawley R. S. Genetic analysis of microtubule motor proteins in Drosophila: a mutation at the ncd locus is a dominant enhancer of nod. Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7165–7169. doi: 10.1073/pnas.88.16.7165. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Komma D. J., Endow S. A. Enhancement of the ncdD microtubule motor mutant by mutants of alpha Tub67C. J Cell Sci. 1997 Jan;110(Pt 2):229–237. doi: 10.1242/jcs.110.2.229. [DOI] [PubMed] [Google Scholar]
- Matthews K. A., Rees D., Kaufman T. C. A functionally specialized alpha-tubulin is required for oocyte meiosis and cleavage mitoses in Drosophila. Development. 1993 Mar;117(3):977–991. doi: 10.1242/dev.117.3.977. [DOI] [PubMed] [Google Scholar]
- Matthies H. J., McDonald H. B., Goldstein L. S., Theurkauf W. E. Anastral meiotic spindle morphogenesis: role of the non-claret disjunctional kinesin-like protein. J Cell Biol. 1996 Jul;134(2):455–464. doi: 10.1083/jcb.134.2.455. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Miki H., Setou M., Kaneshiro K., Hirokawa N. All kinesin superfamily protein, KIF, genes in mouse and human. Proc Natl Acad Sci U S A. 2001 Jun 19;98(13):7004–7011. doi: 10.1073/pnas.111145398. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Myers E. W., Sutton G. G., Delcher A. L., Dew I. M., Fasulo D. P., Flanigan M. J., Kravitz S. A., Mobarry C. M., Reinert K. H., Remington K. A. A whole-genome assembly of Drosophila. Science. 2000 Mar 24;287(5461):2196–2204. doi: 10.1126/science.287.5461.2196. [DOI] [PubMed] [Google Scholar]
- 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]
- Rasooly R. S., New C. M., Zhang P., Hawley R. S., Baker B. S. The lethal(1)TW-6cs mutation of Drosophila melanogaster is a dominant antimorphic allele of nod and is associated with a single base change in the putative ATP-binding domain. Genetics. 1991 Oct;129(2):409–422. doi: 10.1093/genetics/129.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rasooly R. S., Zhang P., Tibolla A. K., Hawley R. S. A structure-function analysis of NOD, a kinesin-like protein from Drosophila melanogaster. Mol Gen Genet. 1994 Jan;242(2):145–151. doi: 10.1007/BF00391007. [DOI] [PubMed] [Google Scholar]
- Riparbelli M. G., Callaini G., Glover D. M. Failure of pronuclear migration and repeated divisions of polar body nuclei associated with MTOC defects in polo eggs of Drosophila. J Cell Sci. 2000 Sep;113(Pt 18):3341–3350. doi: 10.1242/jcs.113.18.3341. [DOI] [PubMed] [Google Scholar]
- Rogers G. C., Chui K. K., Lee E. W., Wedaman K. P., Sharp D. J., Holland G., Morris R. L., Scholey J. M. A kinesin-related protein, KRP(180), positions prometaphase spindle poles during early sea urchin embryonic cell division. J Cell Biol. 2000 Aug 7;150(3):499–512. doi: 10.1083/jcb.150.3.499. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rørth P., Szabo K., Bailey A., Laverty T., Rehm J., Rubin G. M., Weigmann K., Milán M., Benes V., Ansorge W. Systematic gain-of-function genetics in Drosophila. Development. 1998 Mar;125(6):1049–1057. doi: 10.1242/dev.125.6.1049. [DOI] [PubMed] [Google Scholar]
- Tavosanis G., Llamazares S., Goulielmos G., Gonzalez C. Essential role for gamma-tubulin in the acentriolar female meiotic spindle of Drosophila. EMBO J. 1997 Apr 15;16(8):1809–1819. doi: 10.1093/emboj/16.8.1809. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Vale R. D., Fletterick R. J. The design plan of kinesin motors. Annu Rev Cell Dev Biol. 1997;13:745–777. doi: 10.1146/annurev.cellbio.13.1.745. [DOI] [PubMed] [Google Scholar]
- Walczak C. E., Vernos I., Mitchison T. J., Karsenti E., Heald R. A model for the proposed roles of different microtubule-based motor proteins in establishing spindle bipolarity. 1998 Jul 30-Aug 13Curr Biol. 8(16):903–913. doi: 10.1016/s0960-9822(07)00370-3. [DOI] [PubMed] [Google Scholar]
- Waters J. C., Salmon E. D. Chromosomes take an active role in spindle assembly. Bioessays. 1995 Nov;17(11):911–914. doi: 10.1002/bies.950171102. [DOI] [PubMed] [Google Scholar]
- Williams B. C., Dernburg A. F., Puro J., Nokkala S., Goldberg M. L. The Drosophila kinesin-like protein KLP3A is required for proper behavior of male and female pronuclei at fertilization. Development. 1997 Jun;124(12):2365–2376. doi: 10.1242/dev.124.12.2365. [DOI] [PubMed] [Google Scholar]
- Williams B. C., Riedy M. F., Williams E. V., Gatti M., Goldberg M. L. The Drosophila kinesin-like protein KLP3A is a midbody component required for central spindle assembly and initiation of cytokinesis. J Cell Biol. 1995 May;129(3):709–723. doi: 10.1083/jcb.129.3.709. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yang J. T., Laymon R. A., Goldstein L. S. A three-domain structure of kinesin heavy chain revealed by DNA sequence and microtubule binding analyses. Cell. 1989 Mar 10;56(5):879–889. doi: 10.1016/0092-8674(89)90692-2. [DOI] [PubMed] [Google Scholar]
- Zhang P., Hawley R. S. The genetic analysis of distributive segregation in Drosophila melanogaster. II. Further genetic analysis of the nod locus. Genetics. 1990 May;125(1):115–127. doi: 10.1093/genetics/125.1.115. [DOI] [PMC free article] [PubMed] [Google Scholar]
