Home LiteratureArticle Details
PMID: 15563607 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, P.H.S.

The yeast par-1 homologs kin1 and kin2 show genetic and physical interactions with components of the exocytic machinery.

Molecular biology of the cell ·Vol. 16 ·No. 2 ·2005-02-00 ·Pages 532-49

Elbert M, Rossi G, Brennwald P

Abstract

Kin1 and Kin2 are Saccharomyces cerevisiae counterparts of Par-1, the Caenorhabditis elegans kinase essential for the establishment of polarity in the one cell embryo. Here, we present evidence for a novel link between Kin1, Kin2, and the secretory machinery of the budding yeast. We isolated KIN1 and KIN2 as suppressors of a mutant form of Rho3, a Rho-GTPase acting in polarized trafficking. Genetic analysis suggests that KIN1 and KIN2 act downstream of the Rab-GTPase Sec4, its exchange factor Sec2, and several components of the vesicle tethering complex, the Exocyst. We show that Kin1 and Kin2 physically interact with the t-SNARE Sec9 and the Lgl homologue Sro7, proteins acting at the final stage of exocytosis. Structural analysis of Kin2 reveals that its catalytic activity is essential for its function in the secretory pathway and implicates the conserved 42-amino acid tail at the carboxy terminal of the kinase in autoinhibition. Finally, we find that Kin1 and Kin2 induce phosphorylation of t-SNARE Sec9 in vivo and stimulate its release from the plasma membrane. In summary, we report the finding that yeast Par-1 counterparts are associated with and regulate the function of the exocytic apparatus via phosphorylation of Sec9.

MeSH Terms
Amino Acid Sequence Amino Acid Substitution Catalytic Domain Cell Fractionation Conserved Sequence Exocytosis Fungal Proteins Glutathione Transferase/metabolism Membrane Proteins Methionine/metabolism Molecular Sequence Data Phosphoproteins/chemistry,genetics,isolation & purification,metabolism Point Mutation Precipitin Tests Protein Binding Protein Serine-Threonine Kinases/chemistry,genetics,isolation & purification,metabolism Protein Structure, Tertiary Recombinant Fusion Proteins/isolation & purification,metabolism Saccharomyces cerevisiae/genetics,growth & development,metabolism Saccharomyces cerevisiae Proteins/chemistry,genetics,isolation & purification,metabolism Subcellular Fractions Two-Hybrid System Techniques
Chemicals
Fungal Proteins Membrane Proteins Phosphoproteins Recombinant Fusion Proteins Saccharomyces cerevisiae Proteins Methionine Glutathione Transferase KIN1 protein, S cerevisiae KIN2 protein, S cerevisiae Protein Serine-Threonine Kinases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Elbert Maya
Department of Cell and Developmental Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
Rossi Guendalina
Brennwald Patrick
References (55)
55 references, click to expand
  1. Homologs of the synaptobrevin/VAMP family of synaptic vesicle proteins function on the late secretory pathway in S. cerevisiae.
    Cell. 1993 Sep 10;74(5):855-61 PMID: 8374953
  2. IgE receptor type I-dependent regulation of a Rab3D-associated kinase: a possible link in the calcium-dependent assembly of SNARE complexes.
    J Biol Chem. 2001 Nov 16;276(46):42893-900 PMID: 11555639
  3. Insights into autoregulation from the crystal structure of twitchin kinase.
    Nature. 1994 Jun 16;369(6481):581-4 PMID: 8202162
  4. Characterization of the KIN2 gene product in Saccharomyces cerevisiae and comparison between the kinase activities of p145KIN1 and p145KIN2.
    Yeast. 1994 Jan;10(1):113-24 PMID: 8203145
  5. KIN1 and KIN2 protein kinases localize to the cytoplasmic face of the yeast plasma membrane.
    Exp Cell Res. 1994 Jul;213(1):93-9 PMID: 8020610
  6. Sec9 is a SNAP-25-like component of a yeast SNARE complex that may be the effector of Sec4 function in exocytosis.
    Cell. 1994 Oct 21;79(2):245-58 PMID: 7954793
  7. Microtubule-associated protein/microtubule affinity-regulating kinase (p110mark). A novel protein kinase that regulates tau-microtubule interactions and dynamic instability by phosphorylation at the Alzheimer-specific site serine 262.
    J Biol Chem. 1995 Mar 31;270(13):7679-88 PMID: 7706316
  8. Sec6, Sec8, and Sec15 are components of a multisubunit complex which localizes to small bud tips in Saccharomyces cerevisiae.
    J Cell Biol. 1995 Jul;130(2):299-312 PMID: 7615633
  9. Genetic analysis of the Saccharomyces cerevisiae RHO3 gene, encoding a rho-type small GTPase, provides evidence for a role in bud formation.
    Genetics. 1996 Feb;142(2):359-69 PMID: 8852836
  10. Yeast Cdc42 functions at a late step in exocytosis, specifically during polarized growth of the emerging bud.
    J Cell Biol. 2001 Nov 12;155(4):581-92 PMID: 11706050
  11. The fusome and microtubules enrich Par-1 in the oocyte, where it effects polarization in conjunction with Par-3, BicD, Egl, and dynein.
    Curr Biol. 2002 Sep 3;12(17):1524-8 PMID: 12225669
  12. Protein kinase C-dependent phosphorylation of synaptosome-associated protein of 25 kDa at Ser187 potentiates vesicle recruitment.
    J Neurosci. 2002 Nov 1;22(21):9278-86 PMID: 12417653
  13. Ca2+-dependent phosphorylation of syntaxin-1A by the death-associated protein (DAP) kinase regulates its interaction with Munc18.
    J Biol Chem. 2003 Jul 11;278(28):26265-74 PMID: 12730201
  14. The role of PAR-1 in regulating the polarised microtubule cytoskeleton in the Drosophila follicular epithelium.
    Development. 2003 Sep;130(17):3965-75 PMID: 12874119
  15. Phosphorylation of SNAP-23 in activated human platelets.
    J Biol Chem. 2003 Nov 7;278(45):44369-76 PMID: 12930825
  16. Drosophila PAR-1 and 14-3-3 inhibit Bazooka/PAR-3 to establish complementary cortical domains in polarized cells.
    Cell. 2003 Dec 12;115(6):691-704 PMID: 14675534
  17. Regulation of releasable vesicle pool sizes by protein kinase A-dependent phosphorylation of SNAP-25.
    Neuron. 2004 Feb 5;41(3):417-29 PMID: 14766180
  18. Mammalian PAR-1 determines epithelial lumen polarity by organizing the microtubule cytoskeleton.
    J Cell Biol. 2004 Mar 1;164(5):717-27 PMID: 14981097
  19. Structure and expression of the SNF1 gene of Saccharomyces cerevisiae.
    Mol Cell Biol. 1984 Jan;4(1):54-60 PMID: 6366513
  20. Two yeast genes that encode unusual protein kinases.
    Proc Natl Acad Sci U S A. 1987 Sep;84(17):6035-9 PMID: 2957690
  21. Identification of genes required for cytoplasmic localization in early C. elegans embryos.
    Cell. 1988 Feb 12;52(3):311-20 PMID: 3345562
  22. The protein kinase family: conserved features and deduced phylogeny of the catalytic domains.
    Science. 1988 Jul 1;241(4861):42-52 PMID: 3291115
  23. A putative protein kinase gene (kin1+) is important for growth polarity in Schizosaccharomyces pombe.
    Proc Natl Acad Sci U S A. 1990 Nov;87(21):8272-6 PMID: 2236039
  24. High-efficiency transformation of yeast by electroporation.
    Methods Enzymol. 1991;194:182-7 PMID: 2005786
  25. The product of the KIN1 locus in Saccharomyces cerevisiae is a serine/threonine-specific protein kinase.
    Yeast. 1991 Apr;7(3):219-28 PMID: 1652871
  26. MARK, a novel family of protein kinases that phosphorylate microtubule-associated proteins and trigger microtubule disruption.
    Cell. 1997 Apr 18;89(2):297-308 PMID: 9108484
  27. Analysis of a yeast SNARE complex reveals remarkable similarity to the neuronal SNARE complex and a novel function for the C terminus of the SNAP-25 homolog, Sec9.
    J Biol Chem. 1997 Jun 27;272(26):16610-7 PMID: 9195974
  28. Sec2p mediates nucleotide exchange on Sec4p and is involved in polarized delivery of post-Golgi vesicles.
    J Cell Biol. 1997 Jun 30;137(7):1495-509 PMID: 9199166
  29. Mammalian homologues of C. elegans PAR-1 are asymmetrically localized in epithelial cells and may influence their polarity.
    Curr Biol. 1997 Aug 1;7(8):603-6 PMID: 9259552
  30. Multiple regulatory domains on the Byr2 protein kinase.
    Mol Cell Biol. 1997 Oct;17(10):5876-87 PMID: 9315645
  31. Formation of a yeast SNARE complex is accompanied by significant structural changes.
    FEBS Lett. 1997 Sep 22;415(1):49-55 PMID: 9326367
  32. Subunit structure of the mammalian exocyst complex.
    Proc Natl Acad Sci U S A. 1997 Dec 23;94(26):14438-43 PMID: 9405631
  33. Binary interactions of the SNARE proteins syntaxin-4, SNAP23, and VAMP-2 and their regulation by phosphorylation.
    Biochemistry. 1998 Aug 4;37(31):11089-96 PMID: 9693005
  34. Genetic requirements for PIE-1 localization and inhibition of gene expression in the embryonic germ lineage of Caenorhabditis elegans.
    Dev Biol. 1998 Aug 15;200(2):212-24 PMID: 9705228
  35. Genetic evidence for Pak1 autoinhibition and its release by Cdc42.
    Mol Cell Biol. 1999 Jan;19(1):602-11 PMID: 9858584
  36. Differential phosphorylation of syntaxin and synaptosome-associated protein of 25 kDa (SNAP-25) isoforms.
    J Neurochem. 1999 Feb;72(2):614-24 PMID: 9930733
  37. The exocyst is an effector for Sec4p, targeting secretory vesicles to sites of exocytosis.
    EMBO J. 1999 Feb 15;18(4):1071-80 PMID: 10022848
  38. Rho3 of Saccharomyces cerevisiae, which regulates the actin cytoskeleton and exocytosis, is a GTPase which interacts with Myo2 and Exo70.
    Mol Cell Biol. 1999 May;19(5):3580-7 PMID: 10207081
  39. Yeast homologues of tomosyn and lethal giant larvae function in exocytosis and are associated with the plasma membrane SNARE, Sec9.
    J Cell Biol. 1999 Jul 12;146(1):125-40 PMID: 10402465
  40. Sec1p binds to SNARE complexes and concentrates at sites of secretion.
    J Cell Biol. 1999 Jul 26;146(2):333-44 PMID: 10427089
  41. Control of the terminal step of intracellular membrane fusion by protein phosphatase 1.
    Science. 1999 Aug 13;285(5430):1084-7 PMID: 10446058
  42. Phosphorylation of SNAP-23 by the novel kinase SNAK regulates t-SNARE complex assembly.
    Mol Biol Cell. 1999 Dec;10(12):4033-41 PMID: 10588641
  43. The Rho GTPase Rho3 has a direct role in exocytosis that is distinct from its role in actin polarity.
    Mol Biol Cell. 1999 Dec;10(12):4121-33 PMID: 10588647
  44. Changes of synaptotagmin interaction with t-SNARE proteins in vitro after calcium/calmodulin-dependent phosphorylation.
    J Neurochem. 2000 Jan;74(1):209-21 PMID: 10617122
  45. Nerve growth factor-induced phosphorylation of SNAP-25 in PC12 cells: a possible involvement in the regulation of SNAP-25 localization.
    J Neurochem. 2000 May;74(5):2058-66 PMID: 10800949
  46. The Drosophila homolog of C. elegans PAR-1 organizes the oocyte cytoskeleton and directs oskar mRNA localization to the posterior pole.
    Cell. 2000 May 12;101(4):377-88 PMID: 10830165
  47. A Drosophila melanogaster homologue of Caenorhabditis elegans par-1 acts at an early step in embryonic-axis formation.
    Nat Cell Biol. 2000 Jul;2(7):458-60 PMID: 10878812
  48. Protein kinase C phosphorylation of syntaxin 4 in thrombin-activated human platelets.
    J Biol Chem. 2000 Aug 18;275(33):25286-91 PMID: 10856305
  49. Mechanisms of synaptic vesicle exocytosis.
    Annu Rev Cell Dev Biol. 2000;16:19-49 PMID: 11031229
  50. t-SNARE dephosphorylation promotes SNARE assembly and exocytosis in yeast.
    EMBO J. 2001 Feb 1;20(3):411-21 PMID: 11157748
  51. PAR-1 is required for the maintenance of oocyte fate in Drosophila.
    Development. 2001 Apr;128(7):1201-9 PMID: 11245586
  52. Drosophila par-1 is required for oocyte differentiation and microtubule organization.
    Curr Biol. 2001 Jan 23;11(2):75-87 PMID: 11231123
  53. Intermolecular and intramolecular interactions regulate catalytic activity of myotonic dystrophy kinase-related Cdc42-binding kinase alpha.
    Mol Cell Biol. 2001 Apr;21(8):2767-78 PMID: 11283256
  54. PAR-1 is a Dishevelled-associated kinase and a positive regulator of Wnt signalling.
    Nat Cell Biol. 2001 Jul;3(7):628-36 PMID: 11433294
  55. Yeast syntaxins Sso1p and Sso2p belong to a family of related membrane proteins that function in vesicular transport.
    EMBO J. 1993 Nov;12(11):4095-104 PMID: 8223426
Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.e04-07-0549
Published
2005-02-00
Epub
2004-00-24
Pages
532-49
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC545889
Subset
IM
Grants
NIGMS NIH HHS · R01 GM054712 · United States
NEI NIH HHS · T32 EY007138 · United States
NIGMS NIH HHS · GM-54712 · United States
NEI NIH HHS · T32EY007138 · United States
External Links
PubMed source DOI: 10.1091/mbc.e04-07-0549 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: [email protected]

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