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PMID: 10508861 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Cytoplasmic dynein is required for distinct aspects of MTOC positioning, including centrosome separation, in the one cell stage Caenorhabditis elegans embryo.

The Journal of cell biology ·Vol. 147 ·No. 1 ·1999-10-04 ·Pages 135-50

Gönczy P, Pichler S, Kirkham M, Hyman AA

Abstract

We have investigated the role of cytoplasmic dynein in microtubule organizing center (MTOC) positioning using RNA-mediated interference (RNAi) in Caenorhabditis elegans to deplete the product of the dynein heavy chain gene dhc-1. Analysis with time-lapse differential interference contrast microscopy and indirect immunofluorescence revealed that pronuclear migration and centrosome separation failed in one cell stage dhc-1 (RNAi) embryos. These phenotypes were also observed when the dynactin components p50/dynamitin or p150(Glued) were depleted with RNAi. Moreover, in 15% of dhc-1 (RNAi) embryos, centrosomes failed to remain in proximity of the male pronucleus. When dynein heavy chain function was diminished only partially with RNAi, centrosome separation took place, but orientation of the mitotic spindle was defective. Therefore, cytoplasmic dynein is required for multiple aspects of MTOC positioning in the one cell stage C. elegans embryo. In conjunction with our observation of cytoplasmic dynein distribution at the periphery of nuclei, these results lead us to propose a mechanism in which cytoplasmic dynein anchored on the nucleus drives centrosome separation.

MeSH Terms
Amino Acid Sequence Animals Antibodies/immunology Biological Transport Caenorhabditis elegans/cytology,drug effects,embryology,metabolism Cell Nucleus/metabolism Centrosome/metabolism Cytoplasm/metabolism Cytoplasmic Granules/metabolism Dynactin Complex Dyneins/chemistry,genetics,immunology,metabolism Egg Yolk/cytology Embryo, Nonmammalian/cytology,drug effects,metabolism Female Gene Silencing/drug effects Helminth Proteins/genetics,metabolism Male Microtubule-Associated Proteins/genetics,metabolism Models, Biological Molecular Sequence Data RNA, Double-Stranded/genetics,pharmacology Spindle Apparatus/metabolism Time Factors
Chemicals
Antibodies Dynactin Complex Helminth Proteins Microtubule-Associated Proteins RNA, Double-Stranded Dyneins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Gönczy P
European Molecular Biology Laboratory, Heidelberg, D-69117 Germany. gonczy@embl-heidelberg.de
Pichler S
Kirkham M
Hyman A A
References (49)
49 references, click to expand
  1. Kinesin-related KIP3 of Saccharomyces cerevisiae is required for a distinct step in nuclear migration.
    J Cell Biol. 1997 Sep 8;138(5):1023-40 PMID: 9281581
  2. Cortical domains and the mechanisms of asymmetric cell division.
    Trends Cell Biol. 1996 Oct;6(10):382-7 PMID: 15157520
  3. Cortical and cytoplasmic flow polarity in early embryonic cells of Caenorhabditis elegans.
    J Cell Biol. 1993 Jun;121(6):1343-55 PMID: 8509454
  4. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans.
    Nature. 1998 Feb 19;391(6669):806-11 PMID: 9486653
  5. The force-producing mechanism for centrosome separation during spindle formation in vertebrates is intrinsic to each aster.
    J Cell Biol. 1993 Jul;122(2):361-72 PMID: 8320259
  6. Localization of the kinesin-like protein Xklp2 to spindle poles requires a leucine zipper, a microtubule-associated protein, and dynein.
    J Cell Biol. 1998 Nov 2;143(3):673-85 PMID: 9813089
  7. ACT3: a putative centractin homologue in S. cerevisiae is required for proper orientation of the mitotic spindle.
    J Cell Biol. 1994 Oct;127(1):129-38 PMID: 7929558
  8. Cytoplasmic dynein is required for normal nuclear segregation in yeast.
    Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):11172-6 PMID: 8248224
  9. The embryonic cell lineage of the nematode Caenorhabditis elegans.
    Dev Biol. 1983 Nov;100(1):64-119 PMID: 6684600
  10. Mitotic spindle positioning in Saccharomyces cerevisiae is accomplished by antagonistically acting microtubule motor proteins.
    J Cell Biol. 1997 Sep 8;138(5):1041-53 PMID: 9281582
  11. Determination of cell division axes in the early embryogenesis of Caenorhabditis elegans.
    J Cell Biol. 1987 Nov;105(5):2123-35 PMID: 3680373
  12. The dynactin complex is required for cleavage plane specification in early Caenorhabditis elegans embryos.
    Curr Biol. 1998 Oct 8;8(20):1110-6 PMID: 9778526
  13. Cytoplasmic dynein is localized to kinetochores during mitosis.
    Nature. 1990 May 17;345(6272):263-5 PMID: 2139717
  14. Centrosome movement in the early divisions of Caenorhabditis elegans: a cortical site determining centrosome position.
    J Cell Biol. 1989 Sep;109(3):1185-93 PMID: 2768338
  15. Astral microtubule dynamics in yeast: a microtubule-based searching mechanism for spindle orientation and nuclear migration into the bud.
    J Cell Biol. 1997 Nov 17;139(4):985-94 PMID: 9362516
  16. Distribution of microtubules during centriole separation in rat kangaroo (Potorous) cells.
    Cytobios. 1976;15(57):37-43 PMID: 1001019
  17. Dynein and dynactin are localized to astral microtubules and at cortical sites in mitotic epithelial cells.
    Curr Biol. 1998 Apr 23;8(9):541-4 PMID: 9560347
  18. Kinesin and dynein superfamily proteins in organelle transport and cell division.
    Curr Opin Cell Biol. 1998 Feb;10(1):60-73 PMID: 9484596
  19. Golgi vesiculation and lysosome dispersion in cells lacking cytoplasmic dynein.
    J Cell Biol. 1998 Apr 6;141(1):51-9 PMID: 9531547
  20. Molecular characterization of the 50-kD subunit of dynactin reveals function for the complex in chromosome alignment and spindle organization during mitosis.
    J Cell Biol. 1996 Feb;132(4):617-33 PMID: 8647893
  21. Generation of asymmetry and segregation of germ-line granules in early C. elegans embryos.
    Cell. 1983 Nov;35(1):15-25 PMID: 6684994
  22. Cytoplasmic dynein plays a role in mammalian mitotic spindle formation.
    J Cell Biol. 1993 Nov;123(4):849-58 PMID: 8227145
  23. Localization of cytoplasmic dynein to mitotic spindles and kinetochores.
    Nature. 1990 May 17;345(6272):266-8 PMID: 2139718
  24. Light and electron microscopy of rat kangaroo cells in mitosis. I. Formation and breakdown of the mitotic apparatus.
    Chromosoma. 1973;40(1):43-82 PMID: 4344486
  25. 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):669-79 PMID: 8069915
  26. Actin-related protein 1 and cytoplasmic dynein-based motility - what's the connection?
    Trends Cell Biol. 1996 Jun;6(6):212-5 PMID: 15157458
  27. Asters, poles, and transport properties within spindlelike microtubule arrays.
    Cold Spring Harb Symp Quant Biol. 1982;46 Pt 1:263-83 PMID: 7049529
  28. Mechanisms of nuclear positioning.
    J Cell Sci. 1998 Aug;111 ( Pt 16):2283-95 PMID: 9683624
  29. Dissection of cell division processes in the one cell stage Caenorhabditis elegans embryo by mutational analysis.
    J Cell Biol. 1999 Mar 8;144(5):927-46 PMID: 10085292
  30. Genomic structure of a cytoplasmic dynein heavy chain gene from the nematode Caenorhabditis elegans.
    Cell Motil Cytoskeleton. 1995;32(1):26-36 PMID: 8674131
  31. Movement of nuclei along microtubules in Xenopus egg extracts.
    Curr Biol. 1997 Mar 1;7(3):211-4 PMID: 9395409
  32. Asymmetric cell division.
    Nature. 1998 Apr 23;392(6678):775-8 PMID: 9572136
  33. Organelle movement. Dynactin: portrait of a dynein regulator.
    Curr Biol. 1994 Nov 1;4(11):1000-2 PMID: 7874478
  34. MAP 1C is a microtubule-activated ATPase which translocates microtubules in vitro and has dynein-like properties.
    J Cell Biol. 1987 Sep;105(3):1273-82 PMID: 2958482
  35. Cytoplasmic dynein function is essential in Drosophila melanogaster.
    Genetics. 1996 Mar;142(3):865-78 PMID: 8849893
  36. Segregation of holocentric chromosomes at meiosis in the nematode, Caenorhabditis elegans.
    Chromosome Res. 1993 May;1(1):15-26 PMID: 8143084
  37. DYNEINS: molecular structure and cellular function.
    Annu Rev Cell Biol. 1994;10:339-72 PMID: 7888180
  38. ZYG-9, a Caenorhabditis elegans protein required for microtubule organization and function, is a component of meiotic and mitotic spindle poles.
    J Cell Biol. 1998 Jun 1;141(5):1159-68 PMID: 9606208
  39. Disruption of mitotic spindle orientation in a yeast dynein mutant.
    Proc Natl Acad Sci U S A. 1993 Nov 1;90(21):10096-100 PMID: 8234262
  40. PGL-1, a predicted RNA-binding component of germ granules, is essential for fertility in C. elegans.
    Cell. 1998 Sep 4;94(5):635-45 PMID: 9741628
  41. Transient localized accumulation of actin in Caenorhabditis elegans blastomeres with oriented asymmetric divisions.
    Development. 1994 Aug;120(8):2317-28 PMID: 7925032
  42. Immunolocalization of cytoplasmic dynein to lysosomes in cultured cells.
    J Cell Sci. 1992 Jan;101 ( Pt 1):125-37 PMID: 1349021
  43. Overexpression of the dynamitin (p50) subunit of the dynactin complex disrupts dynein-dependent maintenance of membrane organelle distribution.
    J Cell Biol. 1997 Oct 20;139(2):469-84 PMID: 9334349
  44. Xklp2, a novel Xenopus centrosomal kinesin-like protein required for centrosome separation during mitosis.
    Cell. 1996 Jan 12;84(1):49-59 PMID: 8548825
  45. Targeting of motor proteins.
    Science. 1996 Mar 15;271(5255):1539-44 PMID: 8599110
  46. Cleavage plane specification in C. elegans: how to divide the spoils.
    Cell. 1996 Jan 26;84(2):195-8 PMID: 8565065
  47. Cytoplasmic dynein participates in the centrosomal localization of the Golgi complex.
    J Cell Biol. 1992 Sep;118(6):1333-45 PMID: 1387874
  48. Formation of the first cleavage spindle in nematode embryos.
    Dev Biol. 1984 Jan;101(1):61-72 PMID: 6692980
  49. Mitotic spindle poles are organized by structural and motor proteins in addition to centrosomes.
    J Cell Biol. 1997 Sep 8;138(5):1055-66 PMID: 9281583
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.147.1.135
Email
gonczy@embl-heidelberg.de
Published
1999-10-04
Pages
135-50
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2164971
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.147.1.135 PMC Full Text View journal details
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