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PMID: 9442113 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Kar9p is a novel cortical protein required for cytoplasmic microtubule orientation in yeast.

The Journal of cell biology ·Vol. 140 ·No. 2 ·1998-01-26 ·Pages 377-90

Miller RK, Rose MD

Abstract

kar9 was originally identified as a bilateral karyogamy mutant, in which the two zygotic nuclei remained widely separated and the cytoplasmic microtubules were misoriented (Kurihara, L.J., C.T. Beh, M. Latterich, R. Schekman, and M.D. Rose. 1994. J. Cell Biol. 126:911-923.). We now report a general defect in nuclear migration and microtubule orientation in kar9 mutants. KAR9 encodes a novel 74-kD protein that is not essential for life. The kar9 mitotic defect was similar to mutations in dhc1/dyn1 (dynein heavy chain gene), jnm1, and act5. kar9Delta dhc1Delta, kar9Delta jnm1Delta, and kar9Delta act5Delta double mutants were synthetically lethal, suggesting that these genes function in partially redundant pathways to carry out nuclear migration. A functional GFP-Kar9p fusion protein localized to a single dot at the tip of the shmoo projection. In mitotic cells, GFP-Kar9p localized to a cortical dot with both mother-daughter asymmetry and cell cycle dependence. In small-budded cells through anaphase, GFP-Kar9p was found at the tip of the growing bud. In telophase and G1 unbudded cells, no localization was observed. By indirect immunofluorescence, cytoplasmic microtubules intersected the GFP-Kar9p dot. Nocodazole experiments demonstrated that Kar9p's cortical localization was microtubule independent. We propose that Kar9p is a component of a cortical adaptor complex that orients cytoplasmic microtubules.

MeSH Terms
Amino Acid Sequence Cell Cycle Cell Survival Cytoplasm/metabolism Dyneins/genetics,metabolism Green Fluorescent Proteins Luminescent Proteins/genetics,metabolism Microtubules/physiology Molecular Sequence Data Nuclear Proteins/chemistry,genetics,physiology Recombinant Fusion Proteins/metabolism Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins
Chemicals
KAR9 protein, S cerevisiae Luminescent Proteins Nuclear Proteins Recombinant Fusion Proteins Saccharomyces cerevisiae Proteins Green Fluorescent Proteins Dyneins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Miller R K
Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544, USA.
Rose M D
References (50)
50 references, click to expand
  1. 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
  2. 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
  3. Cytoplasmic dynein and actin-related protein Arp1 are required for normal nuclear distribution in filamentous fungi.
    J Cell Biol. 1994 Oct;127(1):139-149 PMID: 7929559
  4. FACS-optimized mutants of the green fluorescent protein (GFP).
    Gene. 1996;173(1 Spec No):33-8 PMID: 8707053
  5. Spindle dynamics and cell cycle regulation of dynein in the budding yeast, Saccharomyces cerevisiae.
    J Cell Biol. 1995 Aug;130(3):687-700 PMID: 7622568
  6. An alpha tubulin mutation suppresses nuclear migration mutations in Aspergillus nidulans.
    Genetics. 1995 Dec;141(4):1287-98 PMID: 8601474
  7. Nuclear fusion in the yeast Saccharomyces cerevisiae.
    Annu Rev Cell Dev Biol. 1996;12:663-95 PMID: 8970740
  8. BIK1, a protein required for microtubule function during mating and mitosis in Saccharomyces cerevisiae, colocalizes with tubulin.
    J Cell Biol. 1990 Dec;111(6 Pt 1):2573-86 PMID: 2277073
  9. One-step gene disruption in yeast.
    Methods Enzymol. 1983;101:202-11 PMID: 6310324
  10. Two genes required for cell fusion during yeast conjugation: evidence for a pheromone-induced surface protein.
    Mol Cell Biol. 1987 Jul;7(7):2316-28 PMID: 3302672
  11. Predicting coiled coils from protein sequences.
    Science. 1991 May 24;252(5009):1162-4 PMID: 2031185
  12. Nuclear congression and membrane fusion: two distinct events in the yeast karyogamy pathway.
    J Cell Biol. 1994 Aug;126(4):911-23 PMID: 8051211
  13. Two Saccharomyces cerevisiae kinesin-related gene products required for mitotic spindle assembly.
    J Cell Biol. 1992 Jul;118(1):109-20 PMID: 1618897
  14. Behavior of spindles and spindle plaques in the cell cycle and conjugation of Saccharomyces cerevisiae.
    J Bacteriol. 1975 Oct;124(1):511-23 PMID: 1100612
  15. Astral microtubules are not required for anaphase B in Saccharomyces cerevisiae.
    J Cell Biol. 1992 Oct;119(2):379-88 PMID: 1400581
  16. Suppression of a myosin defect by a kinesin-related gene.
    Nature. 1992 Mar 26;356(6367):358-61 PMID: 1549181
  17. Molecular cloning of a ubiquitously distributed microtubule-associated protein with Mr 190,000.
    J Biol Chem. 1990 Aug 15;265(23):13849-55 PMID: 2380192
  18. A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli.
    Gene. 1987;57(2-3):267-72 PMID: 3319781
  19. NudF, a nuclear migration gene in Aspergillus nidulans, is similar to the human LIS-1 gene required for neuronal migration.
    Mol Biol Cell. 1995 Mar;6(3):297-310 PMID: 7612965
  20. Structural basis for the binding of proline-rich peptides to SH3 domains.
    Cell. 1994 Mar 11;76(5):933-45 PMID: 7510218
  21. Improved tools for biological sequence comparison.
    Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444-8 PMID: 3162770
  22. KAR3, a kinesin-related gene required for yeast nuclear fusion.
    Cell. 1990 Mar 23;60(6):1029-41 PMID: 2138512
  23. Actin- and tubulin-dependent functions during Saccharomyces cerevisiae mating projection formation.
    Mol Biol Cell. 1992 Apr;3(4):429-44 PMID: 1498363
  24. A Saccharomyces cerevisiae genomic plasmid bank based on a centromere-containing shuttle vector.
    Gene. 1987;60(2-3):237-43 PMID: 3327750
  25. 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
  26. Physical maps of the six smallest chromosomes of Saccharomyces cerevisiae at a resolution of 2.6 kilobase pairs.
    Genetics. 1993 May;134(1):81-150 PMID: 8514151
  27. Kinesin-related proteins required for assembly of the mitotic spindle.
    J Cell Biol. 1992 Jul;118(1):95-108 PMID: 1618910
  28. 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
  29. The JNM1 gene in the yeast Saccharomyces cerevisiae is required for nuclear migration and spindle orientation during the mitotic cell cycle.
    J Cell Biol. 1994 Apr;125(1):143-58 PMID: 8138567
  30. Micromanipulation studies of the asymmetric positioning of the maturation spindle in Chaetopterus sp. oocytes: I. Anchorage of the spindle to the cortex and migration of a displaced spindle.
    Cell Motil Cytoskeleton. 1988;11(2):83-96 PMID: 3191533
  31. 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
  32. Characterization and localization of the cytoplasmic dynein heavy chain in Aspergillus nidulans.
    Proc Natl Acad Sci U S A. 1995 Oct 10;92(21):9890-4 PMID: 7568239
  33. Nuclear fusion in yeast.
    Annu Rev Microbiol. 1991;45:539-67 PMID: 1741623
  34. 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
  35. A role for the actin cytoskeleton of Saccharomyces cerevisiae in bipolar bud-site selection.
    J Cell Biol. 1997 Jan 13;136(1):111-23 PMID: 9008707
  36. Studies concerning the temporal and genetic control of cell polarity in Saccharomyces cerevisiae.
    J Cell Biol. 1991 Aug;114(3):515-32 PMID: 1860883
  37. Saccharomyces cerevisiae genes required in the absence of the CIN8-encoded spindle motor act in functionally diverse mitotic pathways.
    Mol Biol Cell. 1997 Jun;8(6):1035-50 PMID: 9201714
  38. Role of astral microtubules and actin in spindle orientation and migration in the budding yeast, Saccharomyces cerevisiae.
    J Cell Biol. 1992 Nov;119(3):583-93 PMID: 1400594
  39. Proline-rich sequences that bind to Src homology 3 domains with individual specificities.
    Proc Natl Acad Sci U S A. 1995 Apr 11;92(8):3110-4 PMID: 7536925
  40. Antibodies to the kinesin motor domain and CENP-E inhibit microtubule depolymerization-dependent motion of chromosomes in vitro.
    J Cell Biol. 1995 Jan;128(1-2):107-15 PMID: 7822408
  41. 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
  42. Duplication of spindle plaques and integration of the yeast cell cycle.
    Cold Spring Harb Symp Quant Biol. 1974;38:123-31 PMID: 4598635
  43. The dynamics of chromosome movement in the budding yeast Saccharomyces cerevisiae.
    J Cell Biol. 1989 Dec;109(6 Pt 2):3355-66 PMID: 2689456
  44. Kinetics of spindle pole body separation in budding yeast.
    Proc Natl Acad Sci U S A. 1995 Oct 10;92(21):9707-11 PMID: 7568202
  45. Polewards chromosome movement driven by microtubule depolymerization in vitro.
    Nature. 1988 Feb 11;331(6156):499-504 PMID: 3340202
  46. Functions of microtubules in the Saccharomyces cerevisiae cell cycle.
    J Cell Biol. 1988 Oct;107(4):1409-26 PMID: 3049620
  47. KAR1, a gene required for function of both intranuclear and extranuclear microtubules in yeast.
    Cell. 1987 Mar 27;48(6):1047-60 PMID: 3030557
  48. Yeast Num1p associates with the mother cell cortex during S/G2 phase and affects microtubular functions.
    J Cell Biol. 1995 Nov;131(4):1003-14 PMID: 7490278
  49. The molecular cloning and identification of a gene product specifically required for nuclear movement in Aspergillus nidulans.
    J Cell Biol. 1990 Aug;111(2):543-51 PMID: 2199460
  50. Cloning vectors for the synthesis of epitope-tagged, truncated and chimeric proteins in Saccharomyces cerevisiae.
    Gene. 1994 Jun 24;144(1):63-8 PMID: 7517907
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.140.2.377
Published
1998-01-26
Pages
377-90
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2132572
Subset
IM
Grants
NIGMS NIH HHS · R01 GM037739 · United States
NIGMS NIH HHS · GM-37739 · United States
External Links
PubMed source DOI: 10.1083/jcb.140.2.377 PMC Full Text View journal details
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