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PMID: 11086011 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The fission yeast ran GTPase is required for microtubule integrity.

The Journal of cell biology ·Vol. 151 ·No. 5 ·2000-11-27 ·Pages 1101-11

Fleig U, Salus SS, Karig I, Sazer S

Abstract

The microtubule cytoskeleton plays a pivotal role in cytoplasmic organization, cell division, and the correct transmission of genetic information. In a screen designed to identify fission yeast genes required for chromosome segregation, we identified a strain that carries a point mutation in the SpRan GTPase. Ran is an evolutionarily conserved eukaryotic GTPase that directly participates in nucleocytoplasmic transport and whose loss affects many biological processes. Recently a transport-independent effect of Ran on spindle formation in vitro was demonstrated, but the in vivo relevance of these findings was unclear. Here, we report the characterization of a Schizosaccharomyces pombe Ran GTPase partial loss of function mutant in which nucleocytoplasmic protein transport is normal, but the microtubule cytoskeleton is defective, resulting in chromosome missegregation and abnormal cell shape. These abnormalities are exacerbated by microtubule destabilizing drugs, by loss of the spindle checkpoint protein Mph1p, and by mutations in the spindle pole body component Cut11p, indicating that SpRan influences microtubule integrity. As the SpRan mutant phenotype can be partially suppressed by the presence of extra Mal3p, we suggest that SpRan plays a role in microtubule stability.

MeSH Terms
Active Transport, Cell Nucleus/physiology Alleles Cell Cycle Proteins/genetics,metabolism Cell Nucleus/metabolism Chromosomes, Fungal/metabolism Cytoplasm/metabolism Genes, Lethal/physiology Interphase/physiology Microtubule-Associated Proteins/genetics,metabolism Microtubules/enzymology Mitosis/physiology Mutagenesis/physiology Nucleotides/metabolism Phenotype Schizosaccharomyces/cytology,enzymology,genetics Schizosaccharomyces pombe Proteins Spindle Apparatus/metabolism ran GTP-Binding Protein/genetics,metabolism
Chemicals
Cell Cycle Proteins Cut1 protein, S pombe Mal3 protein, S pombe Microtubule-Associated Proteins Nucleotides Schizosaccharomyces pombe Proteins ran GTP-Binding Protein
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Fleig U
Institut für Mikrobiologie, Heinrich-Heine-Universität Düsseldorf, 40225 Düsseldorf, Germany. fleigu@uni-duesseldorf.de
Salus S S
Karig I
Sazer S
References (57)
57 references, click to expand
  1. Cell cycle specificity of certain antimicrotubular drugs in Schizosaccharomyces pombe.
    J Gen Microbiol. 1982 Jan;128(1):61-71 PMID: 7086393
  2. Large-scale screening of intracellular protein localization in living fission yeast cells by the use of a GFP-fusion genomic DNA library.
    Genes Cells. 2000 Mar;5(3):169-90 PMID: 10759889
  3. Spindle assembly and the art of regulating microtubule dynamics by MAPs and Stathmin/Op18.
    Trends Cell Biol. 2000 Jul;10(7):261-7 PMID: 10856928
  4. The Schizosaccharomyces pombe spindle checkpoint protein mad2p blocks anaphase and genetically interacts with the anaphase-promoting complex.
    Proc Natl Acad Sci U S A. 1997 Jul 22;94(15):7965-70 PMID: 9223296
  5. Generation of GTP-bound Ran by RCC1 is required for chromatin-induced mitotic spindle formation.
    Nature. 1999 Jul 8;400(6740):178-81 PMID: 10408446
  6. Self-organization of microtubule asters induced in Xenopus egg extracts by GTP-bound Ran.
    Science. 1999 May 21;284(5418):1356-8 PMID: 10334990
  7. CDK2 encodes a 33-kDa cyclin A-associated protein kinase and is expressed before CDC2 in the cell cycle.
    Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):2907-11 PMID: 1372993
  8. Perturbations in the spi1p GTPase cycle of Schizosaccharomyces pombe through its GTPase-activating protein and guanine nucleotide exchange factor components result in similar phenotypic consequences.
    Mol Cell Biol. 1996 Nov;16(11):6352-62 PMID: 8887664
  9. imp2, a new component of the actin ring in the fission yeast Schizosaccharomyces pombe.
    J Cell Biol. 1998 Oct 19;143(2):415-27 PMID: 9786952
  10. Regulation of cell polarity by microtubules in fission yeast.
    J Cell Biol. 1998 Jul 27;142(2):457-71 PMID: 9679144
  11. Nucleocytoplasmic transport: the soluble phase.
    Annu Rev Biochem. 1998;67:265-306 PMID: 9759490
  12. The ran GTPase regulates mitotic spindle assembly.
    Curr Biol. 1999 May 6;9(9):481-4 PMID: 10322113
  13. A novel nuclear export signal sensitive to oxidative stress in the fission yeast transcription factor Pap1.
    J Biol Chem. 1999 May 21;274(21):15151-8 PMID: 10329722
  14. Kinesin-related cut7 protein associates with mitotic and meiotic spindles in fission yeast.
    Nature. 1992 Mar 5;356(6364):74-6 PMID: 1538784
  15. Fission yeast cell morphogenesis: identification of new genes and analysis of their role during the cell cycle.
    J Cell Biol. 1995 Dec;131(6 Pt 1):1529-38 PMID: 8522609
  16. Identification of the pleiotropic cell division cycle gene NDA2 as one of two different alpha-tubulin genes in Schizosaccharomyces pombe.
    Cell. 1984 May;37(1):233-42 PMID: 6327053
  17. Chromosome segregation: dual control ensures fidelity.
    Curr Biol. 1999 Jul 29-Aug 12;9(15):R562-4 PMID: 10469560
  18. The fission yeast microtubule cytoskeleton.
    J Cell Sci. 1998 Jun;111 ( Pt 12):1603-12 PMID: 9601091
  19. A fission yeast RCC1-related protein is required for the mitosis to interphase transition.
    EMBO J. 1994 Feb 1;13(3):606-15 PMID: 8313905
  20. A cytokinesis checkpoint requiring the yeast homologue of an APC-binding protein.
    Nature. 1998 Jun 4;393(6684):487-91 PMID: 9624007
  21. Mph1, a member of the Mps1-like family of dual specificity protein kinases, is required for the spindle checkpoint in S. pombe.
    J Cell Sci. 1998 Jun;111 ( Pt 12):1635-47 PMID: 9601094
  22. Fission yeast mal2+ is required for chromosome segregation.
    Mol Cell Biol. 1996 Nov;16(11):6169-77 PMID: 8887647
  23. Genetic instability and darwinian selection in tumours.
    Trends Cell Biol. 1999 Dec;9(12):M57-60 PMID: 10611684
  24. Versatile shuttle vectors and genomic libraries for use with Schizosaccharomyces pombe.
    Gene. 1992 May 1;114(1):59-66 PMID: 1587485
  25. Ran-GTP stabilises microtubule asters and inhibits nuclear assembly in Xenopus egg extracts.
    J Cell Sci. 1999 Jul;112 ( Pt 14):2453-61 PMID: 10381400
  26. Heterologous modules for efficient and versatile PCR-based gene targeting in Schizosaccharomyces pombe.
    Yeast. 1998 Jul;14(10):943-51 PMID: 9717240
  27. The ran decathlon: multiple roles of Ran.
    J Cell Sci. 2000 Apr;113 ( Pt 7):1111-8 PMID: 10704362
  28. Sequential alterations in the nuclear chromatin region during mitosis of the fission yeast Schizosaccharomyces pombe: video fluorescence microscopy of synchronously growing wild-type and cold-sensitive cdc mutants by using a DNA-binding fluorescent probe.
    J Cell Sci. 1981 Dec;52:271-87 PMID: 7334057
  29. Stimulation of microtubule aster formation and spindle assembly by the small GTPase Ran.
    Science. 1999 May 21;284(5418):1359-62 PMID: 10334991
  30. Fission yeast minichromosome loss mutants mis cause lethal aneuploidy and replication abnormality.
    Mol Biol Cell. 1994 Oct;5(10):1145-58 PMID: 7865880
  31. Beyond self-assembly: from microtubules to morphogenesis.
    Cell. 1986 May 9;45(3):329-42 PMID: 3516413
  32. The NDA3 gene of fission yeast encodes beta-tubulin: a cold-sensitive nda3 mutation reversibly blocks spindle formation and chromosome movement in mitosis.
    Cell. 1984 Dec;39(2 Pt 1):349-58 PMID: 6094012
  33. The product of the spindle formation gene sad1+ associates with the fission yeast spindle pole body and is essential for viability.
    J Cell Biol. 1995 May;129(4):1033-47 PMID: 7744953
  34. Structure of the nuclear transport complex karyopherin-beta2-Ran x GppNHp.
    Nature. 1999 May 20;399(6733):230-7 PMID: 10353245
  35. Definition of individual components within the cytoskeleton of Trypanosoma brucei by a library of monoclonal antibodies.
    J Cell Sci. 1989 Jul;93 ( Pt 3):491-500 PMID: 2606940
  36. Regulation of the fission yeast transcription factor Pap1 by oxidative stress: requirement for the nuclear export factor Crm1 (Exportin) and the stress-activated MAP kinase Sty1/Spc1.
    Genes Dev. 1998 May 15;12(10):1453-63 PMID: 9585505
  37. Yeast Bim1p promotes the G1-specific dynamics of microtubules.
    J Cell Biol. 1999 May 31;145(5):993-1007 PMID: 10352017
  38. Premature initiation of mitosis in yeast lacking RCC1 or an interacting GTPase.
    Cell. 1991 Jul 26;66(2):347-60 PMID: 1855255
  39. Complex formation between Mad1p, Bub1p and Bub3p is crucial for spindle checkpoint function.
    Curr Biol. 2000 Jun 1;10(11):675-8 PMID: 10837255
  40. The use of cell division cycle mutants to investigate the control of microtubule distribution in the fission yeast Schizosaccharomyces pombe.
    J Cell Sci. 1988 Mar;89 ( Pt 3):343-57 PMID: 3198697
  41. Microtubule dynamics and tubulin interacting proteins.
    Curr Opin Cell Biol. 2000 Feb;12(1):52-6 PMID: 10679354
  42. Analysis of the Saccharomyces spindle pole by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry.
    J Cell Biol. 1998 May 18;141(4):967-77 PMID: 9585415
  43. The spindle checkpoint: two transitions, two pathways.
    Trends Cell Biol. 2000 Apr;10(4):154-8 PMID: 10740270
  44. Mal3, the fission yeast homologue of the human APC-interacting protein EB-1 is required for microtubule integrity and the maintenance of cell form.
    J Cell Biol. 1997 Nov 3;139(3):717-28 PMID: 9348288
  45. A protein that shuttles between the nucleus and the cytoplasm is an important mediator of RNA export.
    Genes Dev. 1996 May 15;10(10):1233-46 PMID: 8675010
  46. The identification of cDNAs that affect the mitosis-to-interphase transition in Schizosaccharomyces pombe, including sbp1, which encodes a spi1p-GTP-binding protein.
    Genetics. 1998 Feb;148(2):645-56 PMID: 9504913
  47. BIM1 encodes a microtubule-binding protein in yeast.
    Mol Biol Cell. 1997 Dec;8(12):2677-91 PMID: 9398684
  48. tea1 and the microtubular cytoskeleton are important for generating global spatial order within the fission yeast cell.
    Cell. 1997 Jun 13;89(6):939-49 PMID: 9200612
  49. Molecular genetic analysis of fission yeast Schizosaccharomyces pombe.
    Methods Enzymol. 1991;194:795-823 PMID: 2005825
  50. cut11(+): A gene required for cell cycle-dependent spindle pole body anchoring in the nuclear envelope and bipolar spindle formation in Schizosaccharomyces pombe.
    Mol Biol Cell. 1998 Oct;9(10):2839-55 PMID: 9763447
  51. The p21 Cdk-interacting protein Cip1 is a potent inhibitor of G1 cyclin-dependent kinases.
    Cell. 1993 Nov 19;75(4):805-16 PMID: 8242751
  52. Structural view of the Ran-Importin beta interaction at 2.3 A resolution.
    Cell. 1999 May 28;97(5):635-46 PMID: 10367892
  53. Transport of macromolecules between the nucleus and the cytoplasm.
    RNA. 1998 Apr;4(4):351-64 PMID: 9630243
  54. When overexpressed, a novel centrosomal protein, RanBPM, causes ectopic microtubule nucleation similar to gamma-tubulin.
    J Cell Biol. 1998 Nov 16;143(4):1041-52 PMID: 9817760
  55. A functional homologue of the RNA1 gene product in Schizosaccharomyces pombe: purification, biochemical characterization, and identification of a leucine-rich repeat motif.
    Mol Biol Cell. 1993 Jun;4(6):569-81 PMID: 8374168
  56. The fission yeast SPB component Cut12 links bipolar spindle formation to mitotic control.
    Genes Dev. 1998 Apr 1;12(7):927-42 PMID: 9531532
  57. The search for the primary function of the Ran GTPase continues.
    Trends Cell Biol. 1996 Mar;6(3):81-5 PMID: 15157482
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.151.5.1101
Email
fleigu@uni-duesseldorf.de
Published
2000-11-27
Pages
1101-11
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2174346
Subset
IM
Grants
NIGMS NIH HHS · GM49119 · United States
Corrections
ErratumIn
-
External Links
PubMed source DOI: 10.1083/jcb.151.5.1101 PMC Full Text View journal details
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