Skip to main content
SpringerLink
Log in

Acidophilic structure and killing mechanism of the Pichia farinosa killer toxin SMKT

  • Chapter
  • First Online:
Microbial Protein Toxins

Part of the book series: Topics in Current Genetics ((TCG,volume 11))

Abstract

SMKT, a killer toxin produced by a halotolerant yeast Pichia farinosa that consists of α and β subunits, is generated from a chromosomally encoded preprotoxin by enzymatic processing. SMKT is only stable under acidic conditions. Under neutral conditions, the subunits easily dissociate, consequently resulting in the aggregation of the α subunit and the concomitant loss of killer activity. The pH sensitive mechanism is discussed based on the crystal structure, circular dichroism, diffusion measurements by NMR, and mass spectrometry data. The SPF1 (sensitivity to the \(\hbox{\emph{\underline{P}.~\underline{\smash{f}}arinosa}}\) killer toxin) gene encoding a novel P-type ATPase was isolated by complementation of SMKT-resistant mutants of Saccharomyces cerevisiae. It is possible that Spf1p and Pmr1p, the Golgi Ca2+ pump, collaborate to maintain ion homeostasis both in the ER and Golgi and contribute to glycosylation and protein trafficking. Possible mechanism of killing by SMKT is discussed in view of the function of Spf1p.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  • 1. Antebi A, Fink GR (1992) The yeast Ca(2+)-ATPase homologue, PMR1, is required for normal Golgi function and localizes in a novel Golgi-like distribution. Mol Biol Cell 3:633-654

    CAS  PubMed  Google Scholar 

  • 2. Antuch W, Guntert P, Wuthrich K (1996) Ancestral bg-crystallin precursor structure in a yeast killer toxin. Nat Struct Biol 3:662-665

    Article  CAS  PubMed  Google Scholar 

  • 3. Axelsen KB, Palmgren MG (1998) Evolution of substrate specificities in the P-type ATPase superfamily. J Mol Evol 46:84-101

    CAS  PubMed  Google Scholar 

  • 4. Ballou L, Cohen RE, Ballou CE (1980) Saccharomyces cerevisiae mutants that make mannoproteins with a truncated carbohydrate outer chain. J Biol Chem 255:5986-5991

    CAS  PubMed  Google Scholar 

  • 5. Belden WJ, Barlowe C (2001) Deletion of yeast p24 genes activates the unfolded protein response. Mol Biol Cell 12:957-969

    CAS  PubMed  Google Scholar 

  • 6. Boone C, Bussey H, Greene D, Thomas DY, Vernet T (1986) Yeast killer toxin: site-directed mutations implicate the precursor protein as the immunity component. Cell 46:105-113

    Article  CAS  PubMed  Google Scholar 

  • 7. Boone C, Sommer SS, Hensel A, Bussey H (1990) Yeast KRE genes provide evidence for a pathway of cell wall b-glucan assembly. J Cell Biol 110:1833-1843

    Article  CAS  PubMed  Google Scholar 

  • 8. Breinig F, Tipper DJ, Schmitt MJ (2002) Kre1p, the plasma membrane receptor for the yeast K1 viral toxin. Cell 108:395-405

    Article  CAS  PubMed  Google Scholar 

  • 9. Bussey H, Boone C, Zhu H, Vernet T, Whiteway M, Thomas DY (1990) Genetic and molecular approaches to synthesis and action of the yeast killer toxin. Experientia 46:193-200

    CAS  PubMed  Google Scholar 

  • 10. Cronin SR, Khoury A, Ferry DK, Hampton RY (2000) Regulation of HMG-CoA reductase degradation requires the P-type ATPase Cod1p/Spf1p. J Cell Biol 148:915-924

    Article  CAS  PubMed  Google Scholar 

  • 11. Cronin SR, Rao R, Hampton RY (2002) Cod1p/Spf1p is a P-type ATPase involved in ER function and Ca2+ homeostasis. J Cell Biol 157:1017-1028

    Article  CAS  PubMed  Google Scholar 

  • 12. de Montigny J, Spehner C, Souciet J, Tekaia F, Dujon B, Wincker P, Artiguenave F, Potier S (2000) Genomic exploration of the hemiascomycetous yeasts: 15. Pichia sorbitophila. FEBS Lett 487:87-90

    Article  PubMed  Google Scholar 

  • 13. Dean N (1995) Yeast glycosylation mutants are sensitive to aminoglycosides. Proc Natl Acad Sci USA 92:1287-1291

    CAS  PubMed  Google Scholar 

  • 14. Deurr G, Strayle J, Plemper R, Elbs S, Klee S, Catty P, Wolf D, Rudolph HK (1998) The medial-Golgi ion pump Pmr1 supplies the yeast secretory pathway with Ca2+ and Mn2+ required for glycosylation, sorting, and endoplasmic reticulum-associated protein degradation. Mol Biol Cell 9:1149-1162

    PubMed  Google Scholar 

  • 15. Dmochowska A, Dignard D, Henning D, Thomas DY, Bussey H (1987) Yeast KEX1 gene encodes a putative protease with a carboxypeptidase B- like function involved in killer toxin and a-factor precursor processing. Cell 50:573-584

    Article  CAS  PubMed  Google Scholar 

  • 16. Facanha AL, Appelgren H, Tabish M, Okorokov L, Ekwall K (2002) The endoplasmic reticulum cation P-type ATPase Cta4p is required for control of cell shape and microtubule dynamics. J Cell Biol 157:1029-1039

    Article  CAS  PubMed  Google Scholar 

  • 17. Fujiki Y, Hubbard AL, Fowler S, Lazarow PB (1982) Isolation of intracellular membranes by means of sodium carbonate treatment: application to endoplasmic reticulum. J Cell Biol 93:97-102

    Article  CAS  PubMed  Google Scholar 

  • 18. Gaynor EC, Mondesert G, Grimme SJ, Reed SI, Orlean P, Emr SD (1999) MCD4 encodes a conserved endoplasmic reticulum membrane protein essential for glycosylphosphatidylinositol anchor synthesis in yeast. Mol Biol Cell 10:627-648

    CAS  PubMed  Google Scholar 

  • 19. Giaever G, Chu AM, Ni L, Connelly C, Riles L, Veronneau S, Dow S, Lucau-Danila A, Anderson K, Andre B, Arkin AP, Astromoff A, El-Bakkoury M, Bangham R, Benito R, Brachat S, Campanaro S, Curtiss M, Davis K, Deutschbauer A, Entian KD, Flaherty P, Foury F, Garfinkel DJ, Gerstein M, Gotte D, Guldener U, Hegemann JH, Hempel S, Herman Z, Jaramillo DF, Kelly DE, Kelly SL, Kotter P, LaBonte D, Lamb DC, Lan N, Liang H, Liao H, Liu L, Luo C, Lussier M, Mao R, Menard P, Ooi SL, Revuelta JL, Roberts CJ, Rose M, Ross-Macdonald P, Scherens B, Schimmack G, Shafer B, Shoemaker DD, Sookhai-Mahadeo S, Storms RK, Strathern JN, Valle G, Voet M, Volckaert G, Wang CY, Ward TR, Wilhelmy J, Winzeler EA, Yang Y, Yen G, Youngman E, Yu K, Bussey H, Boeke JD, Snyder M, Philippsen P, Davis RW, Johnston M (2002) Functional profiling of the Saccharomyces cerevisiae genome. Nature 418:387-391

    Article  CAS  PubMed  Google Scholar 

  • 20. Gu F, Khimani A, Rane SG, Flurkey WH, Bozarth RF, Smith TJ (1995) Structure and function of a virally encoded fungal toxin from Ustilago maydis: a fungal and mammalian Ca2+ channel inhibitor. Structure 3:805-814

    CAS  PubMed  Google Scholar 

  • 21. Harris SL, Na S, Zhu X, Seto-Young D, Perlin DS, Teem JH, Haber JE (1994) Dominant lethal mutations in the plasma membrane H(+)-ATPase gene of Saccharomyces cerevisiae. Proc Natl Acad Sci USA 91:10531-10535

    CAS  PubMed  Google Scholar 

  • 22. Hayman GT, Bolen PL (1991) Linear DNA plasmids of Pichia inositovora are associated with a novel killer toxin activity. Curr Genet 19:389-393

    CAS  PubMed  Google Scholar 

  • 23. Heintel T, Zagorc T, Schmitt MJ (2001) Expression, processing and high level secretion of a virus toxin in fission yeast. Appl Microbiol Biotechnol 56:165-172

    Article  CAS  PubMed  Google Scholar 

  • 24. Jakob CA, Burda P, Roth J, Aebi M (1998) Degradation of misfolded endoplasmic reticulum glycoproteins in Saccharomyces cerevisiae is determined by a specific oligosaccharide structure. J Cell Biol 142:1223-1233

    Article  CAS  PubMed  Google Scholar 

  • 25. Julius D, Brake A, Blair L, Kunisawa R, Thorner J (1984) Isolation of the putative structural gene for the lysine-arginine- cleaving endopeptidase required for processing of yeast prepro-a-factor. Cell 37:1075-1089

    Article  CAS  PubMed  Google Scholar 

  • 26. Kagan BL (1983) Mode of action of yeast killer toxins: channel formation in lipid bilayer membranes. Nature 302:709-711

    CAS  PubMed  Google Scholar 

  • 27. Kashiwagi T, Kunishima N, Suzuki C, Tsuchiya F, Nikkuni S, Arata Y, Morikawa K (1997) The novel acidophilic structure of the killer toxin from halotolerant yeast demonstrates remarkable folding similarity with a fungal killer toxin. Structure 5:81-94

    Article  CAS  PubMed  Google Scholar 

  • 28. Kashiwagi T, Yamada N, Hirayama K, Suzuki C, Kashiwagi Y, Tsuchiya F, Arata Y, Kunishima N, Morikawa K (2000) An electrospray-ionization mass spectrometry analysis of the pH-dependent dissociation and denaturation processes of a heterodimeric protein. J Am Soc Mass Spectrom 11:54-61

    Article  CAS  PubMed  Google Scholar 

  • 29. Knop M, Hauser N, Wolf DH (1996) N-Glycosylation affects endoplasmic reticulum degradation of a mutated derivative of carboxypeptidase yscY in yeast. Yeast 12:1229-1238

    Article  CAS  PubMed  Google Scholar 

  • 30. Kurtzman CP (1998) Pichia EC Hansen emend Kurtzman In: Fell JW (ed) The yeasts, a taxonomic study, 4th ed. edn. Elsevier, pp 273-352

    Google Scholar 

  • 31. Li N, Erman M, Pangborn W, Duax WL, Park CM, Bruenn J, Ghosh D (1999) Structure of Ustilago maydis killer toxin KP6 a-subunit. A multimeric assembly with a central pore. J Biol Chem 274:20425-20431

    Article  CAS  PubMed  Google Scholar 

  • 32. Liu CY, Kaufman RJ (2003) The unfolded protein response. J Cell Sci 116:1861-1862

    Article  CAS  PubMed  Google Scholar 

  • 33. Maresova L, Sychrova H (2003) Physiological characterization of osmotolerant yeast Pichia sorbitophila and comparison with a putative synonym Pichia farinosa. Folia Microbiol (Praha) 48:211-217

    Google Scholar 

  • 34. Meaden P, Hill K, Wagner J, Slipetz D, Sommer SS, Bussey H (1990) The yeast KRE5 gene encodes a probable endoplasmic reticulum protein required for (1-6)-b-D-glucan synthesis and normal cell growth. Mol Cell Biol 10:3013-3019

    CAS  PubMed  Google Scholar 

  • 35. Ogawa N, Mori K (2004) Autoregulation of the HAC1 gene is required for sustained activation of the yeast unfolded protein response. Genes Cells 9:95-104

    Article  CAS  PubMed  Google Scholar 

  • 36. Pfeiffer P, Radler F (1984) Comparison of the killer toxin of several yeasts and the purification of a toxin of type K2. Arch Microbiol 137:357-361

    CAS  PubMed  Google Scholar 

  • 37. Polonelli L, Seguy N, Conti S, Gerloni M, Bertolotti D, Cantelli C, Magliani W, Cailliez JC (1997) Monoclonal yeast killer toxin-like candidacidal anti-idiotypic antibodies. Clin Diagn Lab Immunol 4:142-146

    CAS  PubMed  Google Scholar 

  • 38. Price WS, Tsuchiya F, Suzuki C, Arata Y (1999) Characterization of the solution properties of Pichia farinosa killer toxin using PGSE NMR diffusion measurements. J Biomolec NMR 13:113-117

    Article  CAS  Google Scholar 

  • 39. Ram AF, Wolters A, Ten Hoopen R, Klis FM (1994) A new approach for isolating cell wall mutants in Saccharomyces cerevisiae by screening for hypersensitivity to calcofluor white. Yeast 10:1019-1030

    CAS  PubMed  Google Scholar 

  • 40. Rudolph HK, Antebi A, Fink GR, Buckley CM, Dorman TE, LeVitre J, Davidow LS, Mao JI, Moir DT (1989) The yeast secretory pathway is perturbed by mutations in PMR1, a member of a Ca2+ ATPase family. Cell 58:133-145

    Article  CAS  PubMed  Google Scholar 

  • 41. Sato M, Sato K, Nakano A (2002) Evidence for the intimate relationship between vesicle budding from the ER and the unfolded protein response. Biochem Biophys Res Commun 296:560-567

    Article  CAS  PubMed  Google Scholar 

  • 42. Schmitt M, Breinig F (2002) The viral killer system in yeast: from molecular biology to application. FEMS Microbiol Rev 26:257-276

    Article  CAS  PubMed  Google Scholar 

  • 43. Selkoe DJ (2003) Folding proteins in fatal ways. Nature 426:900-904

    Article  CAS  PubMed  Google Scholar 

  • 44. Shimma Y, Nishikawa A, bin Kassim B, Eto A, Jigami Y (1997) A defect in GTP synthesis affects mannose outer chain elongation in Saccharomyces cerevisiae. Mol Gen Genet 256:469-480

    Article  CAS  PubMed  Google Scholar 

  • 45. Sidrauski C, Walter P (1997) The transmembrane kinase Ire1p is a site-specific endonuclease that initiates mRNA splicing in the unfolded protein response. Cell 90:1031-1039

    Article  CAS  PubMed  Google Scholar 

  • 46. Sorin A, Rosas G, Rao R (1997) PMR1, a Ca2+-ATPase in yeast Golgi, has properties distinct from sarco/endoplasmic reticulum and plasma membrane calcium pumps. J Biol Chem 272:9895-9901

    Article  CAS  PubMed  Google Scholar 

  • 47. Souciet JL, Aigle M, Artiguenave F, Blandin G, Bolotin-Fukuhara M, Bon E, Brottier P, Casaregola S, de Montigny J, Dujon B, Durrens P, Gaillardin C, Lepingle A, Llorente B, Malpertuy A, Neuveglise C, Ozier-Kalogeropoulos O, Potier S, Saurin W, Tekaia F, Toffano-Nioche C, Wesolowski-Louvel M, Wincker P, Weissenbach J (2000) Genomic exploration of the hemiascomycetous yeasts: 1. A set of yeast species for molecular evolution studies. FEBS Lett 487:3-12

    Article  PubMed  Google Scholar 

  • 48. Suzuki C (1999) Secretion of a protoxin post-translationally controlled by NaCl in a halotolerant yeast, Pichia farinosa. Yeast 15:123-131

    Article  CAS  PubMed  Google Scholar 

  • 49. Suzuki C (2001) Immunochemical and mutational analyses of P-type ATPase Spf1p involved in the yeast secretory pathway. Biosci Biotechnol Biochem 65:2405-2411

    Article  CAS  PubMed  Google Scholar 

  • 50. Suzuki C, Ando Y, Machida S (2001) Interaction of SMKT, a killer toxin produced by Pichia farinosa, with the yeast cell membranes. Yeast 18:1471-1478

    Article  CAS  PubMed  Google Scholar 

  • 51. Suzuki C, Kashiwagi T, Hirayama K (2002) Alternative CUG codon usage (Ser for Leu) in Pichia farinosa and the effect of a mutated killer gene in Saccharomyces cerevisiae. Protein Eng 15:251-255

    Article  CAS  PubMed  Google Scholar 

  • 52. Suzuki C, Kashiwagi T, Tsuchiya F, Kunishima N, Morikawa K, Nikkuni S, Arata Y (1997) Circular dichroism analysis of the interaction between the a and b subunits in a killer toxin produced by a halotolerant yeast, Pichia farinosa. Protein Engng 10:99-101

    Article  CAS  Google Scholar 

  • 53. Suzuki C, Kawano M, Kashiwagi T, Arata Y, Kawasumi T, Kashiwagi Y (2000) Lethal effect of the expression of a killer gene SMK1 in Saccharomyces cerevisiae. Protein Eng 13:73-76

    Article  CAS  PubMed  Google Scholar 

  • 54. Suzuki C, Nikkuni S (1989) Purification and properties of the killer toxin produced by a halotolerant yeast, Pichia farinosa. Agric Biol Chem 53:2599-2604

    CAS  Google Scholar 

  • 55. Suzuki C, Nikkuni S (1994) The primary and subunit structure of a novel type killer toxin produced by a halotolerant yeast, Pichia farinosa. J Biol Chem 269:3041-3046

    CAS  PubMed  Google Scholar 

  • 56. Suzuki C, Shimma Y (1999) P-type ATPase spf1 mutants show a novel resistant mechanism for the killer toxin SMKT. Mol Microbiol 32:813-824

    Google Scholar 

  • 57. Suzuki C, Yamada K, Okada N, Nikkuni S (1989) Isolation and characterization of halotolerant killer yeasts from fermented foods. Agric Biol Chem 53:2593-2597

    CAS  Google Scholar 

  • 58. Suzuki C, Yoshida N, Okano E, Kawasumi T, Kashiwagi Y (2003) Cloning and chromosomal mapping of URA3 genes of Pichia farinosa and P. sorbitophila encoding orotidine-5’-phosphate decarboxylase. Yeast 20:905-912

    Article  CAS  PubMed  Google Scholar 

  • 59. Tekaia F, Blandin G, Malpertuy A, Llorente B, Durrens P, Toffano-Nioche C, Ozier-Kalogeropoulos O, Bon E, Gaillardin C, Aigle M, Bolotin-Fukuhara M, Casaregola S, de Montigny J, Lepingle A, Neuveglise C, Potier S, Souciet J, Wesolowski-Louvel M, Dujon B (2000) Genomic exploration of the hemiascomycetous yeasts: 3. Methods and strategies used for sequence analysis and annotation. FEBS Lett 487:17-30

    Article  CAS  PubMed  Google Scholar 

  • 60. Tipper DJ, Harley CA (2002) Yeast genes controlling responses to topogenic signals in a model transmembrane protein. Mol Biol Cell 13:1158-1174

    Article  CAS  PubMed  Google Scholar 

  • 61. Tokunaga M, Kawamura A, Hishinuma F (1989) Expression of pGKL killer 28K subunit in Saccharomyces cerevisiae: identification of 28K subunit as a killer protein. Nucleic Acids Res 17:3435-3446

    CAS  PubMed  Google Scholar 

  • 62. Tong AH, Evangelista M, Parsons AB, Xu H, Bader GD, Page N, Robinson M, Raghibizadeh S, Hogue CW, Bussey H, Andrews B, Tyers M, Boone C (2001) Systematic genetic analysis with ordered arrays of yeast deletion mutants. Science 294:2364-2368

    Article  CAS  PubMed  Google Scholar 

  • 63. Tong AH, Lesage G, Bader GD, Ding H, Xu H, Xin X, Young J, Berriz GF, Brost RL, Chang M, Chen Y, Cheng X, Chua G, Friesen H, Goldberg DS, Haynes J, Humphries C, He G, Hussein S, Ke L, Krogan N, Li Z, Levinson JN, Lu H, Menard P, Munyana C, Parsons AB, Ryan O, Tonikian R, Roberts T, Sdicu AM, Shapiro J, Sheikh B, Suter B, Wong SL, Zhang LV, Zhu H, Burd CG, Munro S, Sander C, Rine J, Greenblatt J, Peter M, Bretscher A, Bell G, Roth FP, Brown GW, Andrews B, Bussey H, Boone C (2004) Global mapping of the yeast genetic interaction network. Science 303:808-813

    Article  CAS  PubMed  Google Scholar 

  • 64. Toyoshima C, Nakasako M, Nomura H, Ogawa H (2000) Crystal structure of the calcium pump of sarcoplasmic reticulum at 2.6 A resolution. Nature 405:647-655

    Article  CAS  PubMed  Google Scholar 

  • 65. Travers KJ, Patil CK, Wodicka L, Lockhart DJ, Weissman JS, Walter P (2000) Functional and genomic analyses reveal an essential coordination between the unfolded protein response and ER-associated degradation. Cell 101:249-258

    Article  CAS  PubMed  Google Scholar 

  • 66. Vashist S, Frank CG, Jakob CA, Ng DT (2002) Two distinctly localized p-type ATPases collaborate to maintain organelle homeostasis required for glycoprotein processing and quality control. Mol Biol Cell 13:3955-3966

    Article  CAS  PubMed  Google Scholar 

  • 67. Weiler F, Rehfeldt K, Bautz F, Schmitt MJ (2002) The Zygosaccharomyces bailii antifungal virus toxin zygocin: cloning and expression in a heterologous fungal host. Mol Microbiol 46:1095-1105

    Article  CAS  PubMed  Google Scholar 

  • 68. Worsham PL, Bolen PL (1990) Killer toxin production in Pichia acaciae is associated with linear DNA plasmids. Curr Genet 18:77-80

    CAS  PubMed  Google Scholar 

  • 69. Yoshida SH, Al-Amodi H, Nakamura T, McInerny CJ, Shimoda C (2003) The Schizosaccharomyces pombe cdt2(+) gene, a target of G1-S phase-specific transcription factor complex DSC1, is required for mitotic and premeiotic DNA replication. Genetics 164:881-893

    CAS  PubMed  Google Scholar 

  • 70. Young TW, Yagiu M (1978) A comparison of the killer character in different yeasts and its classification. Antonie Van Leeuwenhoek 44:59-77

    CAS  PubMed  Google Scholar 

  • 71. Zhu H, Bussey H (1991) Mutational analysis of the functional domains of yeast K1 killer toxin. Mol Cell Biol 11:175-181

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan. Present address: National Institute of Livestock and Grassland Science, 2 Ikenodai, Tsukuba, Ibaraki 305-0901, Japan

    Chise Suzuki

Authors
  1. Chise Suzuki
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Chise Suzuki .

Editor information

Manfred J. Schmitt Raffael Schaffrath

Rights and permissions

Reprints and permissions

About this chapter

Cite this chapter

Suzuki, C. Acidophilic structure and killing mechanism of the Pichia farinosa killer toxin SMKT. In: Schmitt, M.J., Schaffrath, R. (eds) Microbial Protein Toxins. Topics in Current Genetics, vol 11. Springer, Berlin, Heidelberg. https://doi.org/10.1007/b101843

Download citation

Publish with us

Policies and ethics

Search

Navigation