Home LiteratureArticle Details
PMID: 12975346 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The roles of microtubule-based motor proteins in mitosis: comprehensive RNAi analysis in the Drosophila S2 cell line.

The Journal of cell biology ·Vol. 162 ·No. 6 ·2003-09-15 ·Pages 1003-16

Goshima G, Vale RD

Abstract

Kinesins and dyneins play important roles during cell division. Using RNA interference (RNAi) to deplete individual (or combinations of) motors followed by immunofluorescence and time-lapse microscopy, we have examined the mitotic functions of cytoplasmic dynein and all 25 kinesins in Drosophila S2 cells. We show that four kinesins are involved in bipolar spindle assembly, four kinesins are involved in metaphase chromosome alignment, dynein plays a role in the metaphase-to-anaphase transition, and one kinesin is needed for cytokinesis. Functional redundancy and alternative pathways for completing mitosis were observed for many single RNAi knockdowns, and failure to complete mitosis was observed for only three kinesins. As an example, inhibition of two microtubule-depolymerizing kinesins initially produced monopolar spindles with abnormally long microtubules, but cells eventually formed bipolar spindles by an acentrosomal pole-focusing mechanism. From our phenotypic data, we construct a model for the distinct roles of molecular motors during mitosis in a single metazoan cell type.

MeSH Terms
Animals Cell Line DNA-Binding Proteins/genetics,metabolism Drosophila Proteins Drosophila melanogaster Dyneins/antagonists & inhibitors,genetics,metabolism Eukaryotic Cells/metabolism Fluorescent Antibody Technique Kinesins/antagonists & inhibitors,genetics,metabolism Kinetochores/drug effects,metabolism Metaphase/genetics Microtubule-Associated Proteins/genetics,metabolism Microtubules/drug effects,genetics,metabolism Molecular Motor Proteins/antagonists & inhibitors,genetics,metabolism Nuclear Proteins/genetics,metabolism RNA Interference RNA, Small Interfering Recombinant Fusion Proteins Spindle Apparatus/genetics,metabolism
Chemicals
DNA-Binding Proteins Drosophila Proteins Microtubule-Associated Proteins Molecular Motor Proteins Nuclear Proteins RNA, Small Interfering Recombinant Fusion Proteins chromokinesin pav protein, Drosophila Dyneins Kinesins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Goshima Gohta
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94107, USA.
Vale Ronald D
References (48)
48 references, click to expand
  1. Centrosome-independent mitotic spindle formation in vertebrates.
    Curr Biol. 2000 Jan 27;10(2):59-67 PMID: 10662665
  2. Cytoplasmic dynein is required for distinct aspects of MTOC positioning, including centrosome separation, in the one cell stage Caenorhabditis elegans embryo.
    J Cell Biol. 1999 Oct 4;147(1):135-50 PMID: 10508861
  3. Small molecule inhibitor of mitotic spindle bipolarity identified in a phenotype-based screen.
    Science. 1999 Oct 29;286(5441):971-4 PMID: 10542155
  4. Functional coordination of three mitotic motors in Drosophila embryos.
    Mol Biol Cell. 2000 Jan;11(1):241-53 PMID: 10637305
  5. Use of double-stranded RNA interference in Drosophila cell lines to dissect signal transduction pathways.
    Proc Natl Acad Sci U S A. 2000 Jun 6;97(12):6499-503 PMID: 10823906
  6. CENP-meta, an essential kinetochore kinesin required for the maintenance of metaphase chromosome alignment in Drosophila.
    J Cell Biol. 2000 Jul 10;150(1):1-11 PMID: 10893249
  7. Flying through the drosophila cytoskeletal genome.
    J Cell Biol. 2000 Jul 24;150(2):F63-8 PMID: 10908588
  8. The Xenopus chromokinesin Xkid is essential for metaphase chromosome alignment and must be degraded to allow anaphase chromosome movement.
    Cell. 2000 Aug 18;102(4):411-24 PMID: 10966104
  9. Xkid, a chromokinesin required for chromosome alignment on the metaphase plate.
    Cell. 2000 Aug 18;102(4):425-35 PMID: 10966105
  10. Spindle assembly in animal cells.
    Annu Rev Biochem. 2000;69:95-114 PMID: 10966454
  11. Microtubule motors in mitosis.
    Nature. 2000 Sep 7;407(6800):41-7 PMID: 10993066
  12. Cytoplasmic dynein is required for poleward chromosome movement during mitosis in Drosophila embryos.
    Nat Cell Biol. 2000 Dec;2(12):922-30 PMID: 11146657
  13. Cell biology. Do centrosome abnormalities lead to cancer?
    Science. 2001 Apr 20;292(5516):426-9 PMID: 11330289
  14. Nuclear migration: cortical anchors for cytoplasmic dynein.
    Curr Biol. 2001 Apr 17;11(8):R326-9 PMID: 11369225
  15. The chromokinesin Kid is necessary for chromosome arm orientation and oscillation, but not congression, on mitotic spindles.
    J Cell Biol. 2001 Sep 17;154(6):1135-46 PMID: 11564754
  16. Kinetochore dynein: its dynamics and role in the transport of the Rough deal checkpoint protein.
    Nat Cell Biol. 2001 Nov;3(11):1001-7 PMID: 11715021
  17. Molecular motors: from one motor many tails to one motor many tales.
    Trends Cell Biol. 2001 Dec;11(12):477-82 PMID: 11719052
  18. Cytoplasmic dynein/dynactin drives kinetochore protein transport to the spindle poles and has a role in mitotic spindle checkpoint inactivation.
    J Cell Biol. 2001 Dec 24;155(7):1159-72 PMID: 11756470
  19. Two kinesin-like Kin I family proteins in fission yeast regulate the establishment of metaphase and the onset of anaphase A.
    Curr Biol. 2002 Apr 16;12(8):610-21 PMID: 11967147
  20. subito encodes a kinesin-like protein required for meiotic spindle pole formation in Drosophila melanogaster.
    Genetics. 2002 Apr;160(4):1489-501 PMID: 11973304
  21. Molecular dissection of cytokinesis by RNA interference in Drosophila cultured cells.
    Mol Biol Cell. 2002 Jul;13(7):2448-60 PMID: 12134082
  22. The microtubule-destabilizing kinesin XKCM1 regulates microtubule dynamic instability in cells.
    Mol Biol Cell. 2002 Aug;13(8):2718-31 PMID: 12181341
  23. Drosophila EB1 is important for proper assembly, dynamics, and positioning of the mitotic spindle.
    J Cell Biol. 2002 Sep 2;158(5):873-84 PMID: 12213835
  24. Spindle-kinetochore attachment requires the combined action of Kin I-like Klp5/6 and Alp14/Dis1-MAPs in fission yeast.
    EMBO J. 2002 Nov 15;21(22):6015-24 PMID: 12426374
  25. The molecular motor toolbox for intracellular transport.
    Cell. 2003 Feb 21;112(4):467-80 PMID: 12600311
  26. 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):1167-80 PMID: 1740471
  27. Kinesin-related proteins required for structural integrity of the mitotic spindle.
    Cell. 1992 Aug 7;70(3):451-8 PMID: 1643659
  28. The kinesin-like protein KLP61F is essential for mitosis in Drosophila.
    J Cell Biol. 1993 Nov;123(3):665-79 PMID: 8227131
  29. Mutants of the Drosophila ncd microtubule motor protein cause centrosomal and spindle pole defects in mitosis.
    J Cell Sci. 1994 Apr;107 ( Pt 4):859-67 PMID: 8056842
  30. 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):709-23 PMID: 7730406
  31. Self-organization of microtubules into bipolar spindles around artificial chromosomes in Xenopus egg extracts.
    Nature. 1996 Aug 1;382(6590):420-5 PMID: 8684481
  32. XCTK2: a kinesin-related protein that promotes mitotic spindle assembly in Xenopus laevis egg extracts.
    J Cell Biol. 1997 Feb 24;136(4):859-70 PMID: 9049251
  33. Mitochondrial association of a plus end-directed microtubule motor expressed during mitosis in Drosophila.
    J Cell Biol. 1997 Mar 10;136(5):1081-90 PMID: 9060472
  34. Monastral bipolar spindles: implications for dynamic centrosome organization.
    J Cell Sci. 1997 Feb;110 ( Pt 4):451-64 PMID: 9067597
  35. Mutation of a gene for a Drosophila kinesin-like protein, Klp38B, leads to failure of cytokinesis.
    J Cell Sci. 1997 Apr;110 ( Pt 8):945-54 PMID: 9152020
  36. KLP38B: a mitotic kinesin-related protein that binds PP1.
    J Cell Biol. 1997 Jul 28;138(2):395-409 PMID: 9230081
  37. Costal2, a novel kinesin-related protein in the Hedgehog signaling pathway.
    Cell. 1997 Jul 25;90(2):235-45 PMID: 9244298
  38. A chromatin-associated kinesin-related protein required for normal mitotic chromosome segregation in Drosophila.
    J Cell Biol. 1997 Dec 15;139(6):1361-71 PMID: 9396743
  39. A Drosophila kinesin-like protein, Klp38B, functions during meiosis, mitosis, and segmentation.
    Dev Biol. 1997 Nov 15;191(2):284-96 PMID: 9398441
  40. Kinesin and dynein superfamily proteins and the mechanism of organelle transport.
    Science. 1998 Jan 23;279(5350):519-26 PMID: 9438838
  41. pavarotti encodes a kinesin-like protein required to organize the central spindle and contractile ring for cytokinesis.
    Genes Dev. 1998 May 15;12(10):1483-94 PMID: 9585508
  42. Cytokinesis and midzone microtubule organization in Caenorhabditis elegans require the kinesin-like protein ZEN-4.
    Mol Biol Cell. 1998 Aug;9(8):2037-49 PMID: 9693365
  43. A model for the proposed roles of different microtubule-based motor proteins in establishing spindle bipolarity.
    Curr Biol. 1998 Jul 30-Aug 13;8(16):903-13 PMID: 9707401
  44. A nematode kinesin required for cleavage furrow advancement.
    Curr Biol. 1998 Oct 8;8(20):1133-6 PMID: 9778533
  45. The bipolar kinesin, KLP61F, cross-links microtubules within interpolar microtubule bundles of Drosophila embryonic mitotic spindles.
    J Cell Biol. 1999 Jan 11;144(1):125-38 PMID: 9885249
  46. Kin I kinesins are microtubule-destabilizing enzymes.
    Cell. 1999 Jan 8;96(1):69-78 PMID: 9989498
  47. Cytoplasmic dynein is required for the nuclear attachment and migration of centrosomes during mitosis in Drosophila.
    J Cell Biol. 1999 Aug 9;146(3):597-608 PMID: 10444068
  48. Mitotic motors in Saccharomyces cerevisiae.
    Biochim Biophys Acta. 2000 Mar 17;1496(1):99-116 PMID: 10722880
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.200303022
Published
2003-09-15
Pages
1003-16
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2172859
Subset
IM
Grants
NIGMS NIH HHS · R01 GM038499 · United States
External Links
PubMed source DOI: 10.1083/jcb.200303022 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2