1887

Abstract

Summary: A 2.5 kb DNA fragment of the gene was cloned by complementation of the mutations that simultaneously lead to resistance to the phytotoxin syringomycin and sensitivity of growth to high Caconcentrations. Sequencing of this fragment revealed a single open reading frame encoding a polypeptide of 365 amino acids. Four hydrophobic regions each separated by hydrophilic regions were present in the protein. was identical to , which is suggested to encode C-5 sterol desaturase required for ergosterol biosynthesis. The protein product of was identified by Western blot analysis as a protein of 40 kDa in the particulate fraction. Gene disruption experiments demonstrated that elimination of is not lethal, but results in membrane C-5 desaturated sterol deficiencies, resistance to syringomycin and sensitivity to high Ca. The mutant cells had significantly decreased ability for syringomycin binding. The results indicated that C-5 desaturated sterols are involved in the binding of syringomycin to the cell, and the lack of the sterols in the mutant membrane results in sensitivity to high Caand an increased rate of cellular Cainflux.

Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-140-2-353
1994-02-01
2024-04-24
Loading full text...

Full text loading...

/deliver/fulltext/micro/140/2/mic-140-2-353.html?itemId=/content/journal/micro/10.1099/13500872-140-2-353&mimeType=html&fmt=ahah

References

  1. Arthington B. A., Benett L. G., Skatrud P. L., Guynn C. J., Barbuch R. J., Ulbright C. E., Bard M. Bard; Cloning, disruption and sequence of the gene encoding yeast C-5 sterol desaturase. Gene 102:39–44
    [Google Scholar]
  2. Ashman W. H., Barbuch R. J., Ulbright C. E., Jarnett H. W., Bard M. Bard; Cloning and disruption of the yeast C-8 sterol isomerase gene. Lipids 26:628–632
    [Google Scholar]
  3. Bard M. Bard; Biochemical and genetic aspects of nystatin resistance in Saccharomyces cerevisiae. J Bacterial 111:649–657
    [Google Scholar]
  4. Bard M., Lees N. D. Burrows, L S., Kleinhans F. W. 1978; Differences in crystal violet uptake and cation-induced death among yeast sterol mutants. J Bacteriol 135:1146–1148
    [Google Scholar]
  5. Bidwai A. P., Takemoto J. Y. 1987; Bacterial phytotoxin, syringomycin, induces a protein kinase-mediated phosphorylation of red beet plasma membrane polypeptides. Proc Natl Acad Sci USA 84:6755–6759
    [Google Scholar]
  6. Breivik O. N., Owades J. L. 1957; Spectrophotometric semi¬micro determination of ergosterol in yeast. Agric Food Chem 5:360–363
    [Google Scholar]
  7. De Vay J. E., Lukezic F. L., Sinden S. L. English, H., Coplin D. L. 1968; A biocide produced by pathogenic isolates of Pseudomonas syringae and its possible role in the bacterial canker disease of peach trees. Phytopathology 58:95–101
    [Google Scholar]
  8. Fukuchi N., Isogai A., Yamashita S., Suyama K., Takemoto J. Y., Suzuki A. Suzuki; Structure of phytotoxin syringomycin produced by a sugar cane isolate of Pseudomonas syringae pv. syringae. Tetrahedron Eett 31:1589–1592
    [Google Scholar]
  9. Gaber R. F., Copple D. M., Kennedy B. K. Vida, M., Bard M. Bard; The yeast gene ERG6 is required for normal membrane function but is not essential for biosynthesis of the cell-cycle- sparking sterol. Mol Cell Biol 9:3447–3456
    [Google Scholar]
  10. Ito H., Fukuda Y. Murata, K., Kimura A. Kimura; T ransformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168
    [Google Scholar]
  11. Jackson M. R. Nilsson, T., Peterson P. A. 1990; Identification of a consensus motif for retention of transmembrane proteins in the endoplasmic reticulum. EMBO J 9:3153–3161
    [Google Scholar]
  12. Lampen J. O., Arnow P. M., Borowska Z., Laskin A. I. 1962; Location and role of sterols at nystatin-binding sites. J Bacteriol 84:1152–1160
    [Google Scholar]
  13. Latoud C. Peypoux, F., Michel G. Michel; Interaction of iturin A, a lipopeptide antibiotic, with Saccharomyces cerevisiae cells: influence of the sterol membrane composition. Can J Microbiol 36:384–389
    [Google Scholar]
  14. Liu Y., Ishii S. Tokai, M„, Tsutsumi H., Ohki O., Akada R., Tanaka K., Tsuchiya E. Fukui, S., Miyakawa T. Miyakawa; The Saccharomyces cerevisiae genes (CMP1 and CMP 2) encoding calmodulin-binding proteins homologous to the catalytic subunit of mammalian protein phosphatase 2B. Mol Gen Genet 227:52–59
    [Google Scholar]
  15. Ohya Y., Umemoto N., Tanida I., Ohta A. lida, H. , Anraku Y. Anraku; Calcium-sensitive els mutants of Saccharomyces cerevisiae showing a Pet- phenotype are ascribable to defects of vacuolar membrane H+-ATPase activity. J Biol Chem 266:13971–13977
    [Google Scholar]
  16. Osumi T. Nishino, T. , Katsuki H. Katsuki; Studies on the A°-desaturation in ergosterol biosynthesis in yeast. T Biochem 85:819–826
    [Google Scholar]
  17. Rothstein R. j. 1983; One-step gene disruption in yeast. Methods Tzn^ymol 101:202–211
    [Google Scholar]
  18. Sanger F. Nicklen, S. , Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
    [Google Scholar]
  19. Segre A., Bachman R. C., Ballio A., Bossa F., Grgurina I., lacobellis N. S., Pucci P. Simmaco, M. , Takemoto J. Y. 1989; The structure of syringomycin Al, E and G. FEBS Lett 255:27–31
    [Google Scholar]
  20. Shin J., Dunbrack R. L. Jr, Lee, S. , Strominger J. L. 1991; Signals for retention of transmembrane proteins in the endoplasmic reticulum studied with CD4 truncation mutants. Proc Natl Acad Sci USA 88:1918–1922
    [Google Scholar]
  21. Sinden S. L., De Vey J. E., Backman P. A. 1971; Properties of syringomycin, a wide spectrum antibiotic and phytotoxin produced by Pseudomonas syringae, and its role in the bacterial canker disease of peach trees. Physiol Plant Pathol 1:199–213
    [Google Scholar]
  22. Takemoto J. Y., Zhang L., Taguchi N. Tachikawa, T. , Miyakawa T. Miyakawa; Mechanism of action of the phytotoxin syringomycin: a resistant mutant of Saccharomyces cerevisiae reveals an involvement of Ca2+ transport. J Gen Microbiol 137:653–659
    [Google Scholar]
  23. Tanaka K., Matsumoto, K. Toh-E A. E; IRA 1, an inhibitory regulator of the Rv4j’-cyclic AMP pathway in Saccharomyces cerevisiae. Mol Cell Biol 9:757–768
    [Google Scholar]
  24. Woods R. A. 1971; Nystatin -resistant mutants of yeast: alter¬ations in sterol content. J Bacteriol 108:69–73
    [Google Scholar]
  25. Zhang, L. Takemoto J. Y. 1986; Mechanism of action of Pseudomonas syringae phytotoxin, syringomycin. Interaction with the plasma membrane of wild-type and respiratory-deficient strains of Saccharomyces cerevisiae. Biochim Biophys Acta 861:201–204
    [Google Scholar]
  26. Zhang, L. Takemoto J. Y. 1987; Syringomycin stimulation of potassium efflux by yeast cells. Biochim Biophys Acta 987:171–175
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/13500872-140-2-353
Loading
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error