Skip to main content
Log in

Immunity and resistance to the KP6 toxin of Ustilago maydis

  • Original Articles
  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

The KP6 toxin of Ustilago maydis, encoded by segmented double-stranded (ds) RNA viruses, is lethal to sensitive strains of the same species and related species. The toxin consists of two polypeptides, α and β, synthesized as a single preprotoxin, which are not covalently linked. Neither polypeptide alone is toxic, but killer activity can be restored by in vitro and in vivo complementation. Killer-secreting strains are resistant to the toxin they produce. Resistance is conferred by a single recessive nuclear gene. This study describes a search for cytoplasmic factors that may confer resistance, also referred to as immunity. The approaches used to detect cytoplasmic immunity included transmission of dsRNA and transmission of virus particles to sensitive cells by cytoduction, cytoplasmic mixing in diploids and infection with viruses. An alternative approach was also used to express cloned cDNAs of the KP6 toxin-encoding dsRNA and of the α and β polypeptides. The results indicated that no immunity to KP6 can be detected. While KP6, α and β polypeptides were expressed by resistant cells, neither KP6 nor β were expressed in sensitive strains. The α polypeptide was expressed in sensitive cells, but it did not confer immunity. These results suggest that neither the preprotoxin nor the α or β polypeptides confer immunity and thus β may be the toxic component of the binary toxin.

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

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Ben-Zvi BS, Koltin Y, Mevarech M, Tamarkin A (1984) RNA polymerase activity in virions from Ustilago maydis. Mol Cell Biol 4:188–194

    Google Scholar 

  • 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 

  • Bostian KA, Eillott Q, Bussey H, Burn V, Smith A, Tipper DJ (1984) Sequence of the preprotoxin dsRNA gene type 1 killer yeast: multiple processing events produce a two-component toxin. Cell 36:741–751

    Article  CAS  PubMed  Google Scholar 

  • Bussey H, Saville D, Greene DJ, Bostian KA (1983) Secretion of Saccharomyces cerevisiae killer toxin: processing of the glycosylated precursor. Mol Cell Biol 3:1362–1370

    CAS  PubMed  Google Scholar 

  • 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

    Google Scholar 

  • Chang TS, Banerjee N, Bruenn JA, Held W, Peery T, Koltin Y (1988) A very small viral dsRNA. Virus Genes 2:195–206

    Google Scholar 

  • Day PR (1981) Fungal virus populations in corn smut from Connecticut. Mycologia 73:379–391

    CAS  Google Scholar 

  • Diamond ME, Dowhanick J, Nemeroff ME, Pietras DF, Tu CL, Bruenn JA (1989) Overlapping genes in a yeast dsRNA virus. J Virol 63:3983–3990

    Google Scholar 

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

    Google Scholar 

  • El-Sherbeini M, Bostian KA (1987) Viruses in fungi: Infection of yeast with the K1 and K2 killer virus. Proc Natl Acad Sci USA 84:4293–4297

    CAS  PubMed  Google Scholar 

  • Eng W-K, Faucette L, Johnson RK, Sternglanz R (1988) Evidence that the DNA topoisomerase I is necessary for the cytotoxic effects of camptothecin. Mol Pharmacol 34:755–760

    Google Scholar 

  • Field LJ, Bruenn JA, Chang TH, Pinhasi O, Koltin Y (1983) Two Ustilago maydis viral dsRNA of different size code for the same product. Nucleic Acids Res 11:2765–2778

    Google Scholar 

  • Finkler A, Koltin Y, Barash I, Sneh B, Pozniak D (1985) Isolation of a virus from virulent strains of Rhizoctonia solani. J Gen Virol 66:1221–1232

    Google Scholar 

  • Fujimura T, Wickner RB (1988) Gene overlap results in a viral protein having an RNA binding domain and a major coat protein domain. Cell 55:663–671

    Google Scholar 

  • Ginzberg I (1990) The Ustilago maydis killer system: The organization of the toxin encoding genes and characterization of their products. PhD thesis, Tel-Aviv University, Israel

    Google Scholar 

  • Holiday R (1961) The genetics of U. maydis. Genet Res 2:204–230

    Google Scholar 

  • Kinal H, Tao J, Bruenn JA (1991) An expression vector for the phytopahtogenic fungus, Ustilago maydis. Gene 98:129–134

    Article  CAS  PubMed  Google Scholar 

  • Koltin Y (1988) The killer system of Ustilago maydis: Secreted polypeptides encoded by viruses. In: Koltin Y, Leibowitz M (eds) Viruses of Fungi and Simple Eukaryotes. Marcel Dekker, New York, pp 209–242

    Google Scholar 

  • Koltin Y, Day PR (1976) Inheritance of killer phenotype and double-stranded RNA in Ustilago maydis. Proc Natl Acad Sci USA 73:594–598

    CAS  PubMed  Google Scholar 

  • Koltin Y, Kandel JS (1978) Killer phenomenon in Ustilago maydis: The organization of the viral genome. Genetics 88:267–276

    Google Scholar 

  • Koltin Y, Mayer I, Steinlauf R (1978) Killer phenomenon in Ustilago maydis: mapping viral functions. Mol Gen Genet 166:181–186

    Google Scholar 

  • Koltin Y, Levine R, Peery T (1980) Assignment of functions to segments of the dsRNA genome of Ustilago maydis. Mol Gen Genet 178:173–178

    CAS  PubMed  Google Scholar 

  • Lolle SJ, Skipper N, Bussey H, Thomas DY (1984) The expression of cDNA clones of yeast M1 double-stranded RNA in yeast confers both killer and immunity phenotypes. EMBO J 3:1383–1387

    CAS  PubMed  Google Scholar 

  • McKoen F (1991) When worlds collide: Immunosuppressant meet protein phosphatases. Cell 66:823–826

    Google Scholar 

  • Peery T, Koltin Y, Tamarkin A (1982) Mapping the immunity function of the Ustilago maydis P1 virus. Plasmid 7:52–58

    CAS  PubMed  Google Scholar 

  • Peery T, Shabat-Brand T, Steinlauf R, Koltin Y, Bruenn J (1987) Virus-encoded toxin of Ustilago maydis: two polypeptides are essential for activity. Mol Cell Biol 7:470–477

    Google Scholar 

  • Puhalla JE (1968) Compatibility reactions on solid medium and interstrain inhibition in Ustilago maydis. Genetics 60:461–474

    CAS  PubMed  Google Scholar 

  • Puhalla JE (1969) The formation of diploids of Ustilago maydis on agar medium. Phytopathology 59:1771–1772

    Google Scholar 

  • Schmitt ME, Brown TA, Trumpower BL (1990) A rapid and simple method for preparation of RNA from Saccharomyces cerevisiae. Nucleic Acids Res 18:3091–3092

    Google Scholar 

  • Seroussi E, Peery T, Ginzberg I, Koltin Y (1989) Detection of killer independent dsRNA plasmids in Ustilago maydis by a simple and rapid method of extraction of dsRNA. Plasmid 21:216–225

    Google Scholar 

  • Smith TL, Leong SA (1990) Isolation and characterization of a Ustilago maydis glyceraldehyde-3-phosphate dehydrogenaseencoding gene. Gene 93:111–117

    Google Scholar 

  • Sturley SL, Elliot Q, LeVitre J, Tipper DJ, Bostian KA (1986) Mapping of functional domains within the Saccharomyces cerevisiae type 1 killer preprotoxin. EMBO J 5:3381–3389

    CAS  PubMed  Google Scholar 

  • Tao J, Ginzberg I, Banerjee N, Held W, Koltin Y, Bruenn JA (1990) Ustilago maydis KP6 toxin: Structure, expression in Saccharomyces cerevisiae, and relationship to other cellular toxins. Mol Cell Biol 10:1373–1381

    CAS  PubMed  Google Scholar 

  • Thomas JO, Kornberg RD (1975) An octamer of histones in chromatin and free in solution. Proc Natl Acad Sci USA 72:2626–2630

    Google Scholar 

  • Tsukuda H, Carleton S, Fotheringham S, Hollman WK (1988) Isolation and characterization of an autonomously replicating sequence from Ustilago maydis. Mol Cell Biol 8:3703–3709

    CAS  PubMed  Google Scholar 

  • Wang J, Holden DW, Leong SA (1988) Gene transfer system for the phytopathogenic fungus Ustilago maydis. Proc Natl Acad Sci USA 5:865–869

    Google Scholar 

  • Ward AC (1990) Single-step purification of shuttle vectors from yeast for high frequency back-transformation into E. coli. Nucleic Acids Res 8:5319

    Google Scholar 

  • Wickner RB (1986) Double-stranded RNA replication in yeast: the killer system. Annu Rev Biochem 55:373–395

    Google Scholar 

  • Widgerson M, Koltin Y (1982) Dual toxin specificities and the exclusion relations among the Ustilago dsRNA viruses. Curr Genet 5:127–136

    Google Scholar 

  • Zhu H, Bussey H (1991) Mutational analysis of the fuctional domains of yeast kl killer toxin. Mol Cell Biol 11:175–181

    Google Scholar 

  • Zhu H, Bussey H, Thomas DY, Gagnon J, Bell AW (1987) Determination of the carboxyl termini of the a and (3 subunits of yeast kl killer toxin. J Biol Chem 262:10728–10732

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by C.P. Hollenberg

Rights and permissions

Reprints and permissions

About this article

Cite this article

Finkler, A., Peery, T., Tao, J. et al. Immunity and resistance to the KP6 toxin of Ustilago maydis . Molec. Gen. Genet. 233, 395–403 (1992). https://doi.org/10.1007/BF00265436

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00265436

Key words

Navigation