Skip to main content
NCBI home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1993 Oct;13(10):6346–6356. doi: 10.1128/mcb.13.10.6346

The yeast KRE9 gene encodes an O glycoprotein involved in cell surface beta-glucan assembly.

J L Brown 1, H Bussey 1
PMCID: PMC364693  PMID: 8413233

Abstract

The yeast KRE9 gene encodes a 30-kDa secretory pathway protein involved in the synthesis of cell wall (1-->6)-beta-glucan. Disruption of KRE9 leads to serious growth impairment and an altered cell wall containing less than 20% of the wild-type amount of (1-->6)-beta-glucan. Analysis of the glucan material remaining in a kre9 delta null mutant indicated a polymer with a reduced average molecular mass. kre9 delta null mutants also displayed several additional cell-wall-related phenotypes, including an aberrant multiply budded morphology, a mating defect, and a failure to form projections in the presence of alpha-factor. Double mutants were generated by crossing kre9 delta strains with strains harboring a null mutation in the KRE1, KRE6, or KRE11 gene, and each of these double mutants was found to be inviable in the SEY6210 background. Similar crosses with null mutations in the KRE5 and SKN1 genes indicated that these double mutants were no more severely affected than kre5 delta or kre9 delta single mutants alone. Antibodies were generated against Kre9p and detected an O glycoprotein of approximately 55 to 60 kDa found in the extracellular medium of a strain overproducing Kre9p.

Full text

PDF
6346

Images in this article

6349Image
on p.6349
6352Image
on p.6352
6353Image
on p.6353
6354Image
on p.6354

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BADIN J., JACKSON C., SCHUBERT M. Improved method for determination of plasma polysaccharides with tryptophan. Proc Soc Exp Biol Med. 1953 Nov;84(2):289–291. [PubMed] [Google Scholar]
  2. Boone C., Sommer S. S., Hensel A., Bussey H. Yeast KRE genes provide evidence for a pathway of cell wall beta-glucan assembly. J Cell Biol. 1990 May;110(5):1833–1843. doi: 10.1083/jcb.110.5.1833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brill S. J., Stillman B. Replication factor-A from Saccharomyces cerevisiae is encoded by three essential genes coordinately expressed at S phase. Genes Dev. 1991 Sep;5(9):1589–1600. doi: 10.1101/gad.5.9.1589. [DOI] [PubMed] [Google Scholar]
  4. Brown J. L., Kossaczka Z., Jiang B., Bussey H. A mutational analysis of killer toxin resistance in Saccharomyces cerevisiae identifies new genes involved in cell wall (1-->6)-beta-glucan synthesis. Genetics. 1993 Apr;133(4):837–849. doi: 10.1093/genetics/133.4.837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bulawa C. E. CSD2, CSD3, and CSD4, genes required for chitin synthesis in Saccharomyces cerevisiae: the CSD2 gene product is related to chitin synthases and to developmentally regulated proteins in Rhizobium species and Xenopus laevis. Mol Cell Biol. 1992 Apr;12(4):1764–1776. doi: 10.1128/mcb.12.4.1764. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bulawa C. E., Slater M., Cabib E., Au-Young J., Sburlati A., Adair W. L., Jr, Robbins P. W. The S. cerevisiae structural gene for chitin synthase is not required for chitin synthesis in vivo. Cell. 1986 Jul 18;46(2):213–225. doi: 10.1016/0092-8674(86)90738-5. [DOI] [PubMed] [Google Scholar]
  7. Bussey H., Sacks W., Galley D., Saville D. Yeast killer plasmid mutations affecting toxin secretion and activity and toxin immunity function. Mol Cell Biol. 1982 Apr;2(4):346–354. doi: 10.1128/mcb.2.4.346. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Costigan C., Gehrung S., Snyder M. A synthetic lethal screen identifies SLK1, a novel protein kinase homolog implicated in yeast cell morphogenesis and cell growth. Mol Cell Biol. 1992 Mar;12(3):1162–1178. doi: 10.1128/mcb.12.3.1162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Drubin D. G. Development of cell polarity in budding yeast. Cell. 1991 Jun 28;65(7):1093–1096. doi: 10.1016/0092-8674(91)90001-f. [DOI] [PubMed] [Google Scholar]
  10. Farkas I., Hardy T. A., Goebl M. G., Roach P. J. Two glycogen synthase isoforms in Saccharomyces cerevisiae are coded by distinct genes that are differentially controlled. J Biol Chem. 1991 Aug 25;266(24):15602–15607. [PubMed] [Google Scholar]
  11. Fleet G. H., Manners D. J. Isolation and composition of an alkali-soluble glucan from the cell walls of Saccharomyces cerevisiae. J Gen Microbiol. 1976 May;94(1):180–192. doi: 10.1099/00221287-94-1-180. [DOI] [PubMed] [Google Scholar]
  12. Friis J., Ottolenghi P. The genetically determined binding of alcian blue by a minor fraction of yeast cell walls. C R Trav Lab Carlsberg. 1970;37(15):327–341. [PubMed] [Google Scholar]
  13. Herscovics A., Orlean P. Glycoprotein biosynthesis in yeast. FASEB J. 1993 Apr 1;7(6):540–550. doi: 10.1096/fasebj.7.6.8472892. [DOI] [PubMed] [Google Scholar]
  14. Hill K., Boone C., Goebl M., Puccia R., Sdicu A. M., Bussey H. Yeast KRE2 defines a new gene family encoding probable secretory proteins, and is required for the correct N-glycosylation of proteins. Genetics. 1992 Feb;130(2):273–283. doi: 10.1093/genetics/130.2.273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Häusler A., Ballou L., Ballou C. E., Robbins P. W. Yeast glycoprotein biosynthesis: MNT1 encodes an alpha-1,2-mannosyltransferase involved in O-glycosylation. Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):6846–6850. doi: 10.1073/pnas.89.15.6846. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  18. Lee K. S., Levin D. E. Dominant mutations in a gene encoding a putative protein kinase (BCK1) bypass the requirement for a Saccharomyces cerevisiae protein kinase C homolog. Mol Cell Biol. 1992 Jan;12(1):172–182. doi: 10.1128/mcb.12.1.172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Manners D. J., Masson A. J., Patterson J. C., Björndal H., Lindberg B. The structure of a beta-(1--6)-D-glucan from yeast cell walls. Biochem J. 1973 Sep;135(1):31–36. doi: 10.1042/bj1350031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Manners D. J., Masson A. J., Patterson J. C. The structure of a beta-(1 leads to 3)-D-glucan from yeast cell walls. Biochem J. 1973 Sep;135(1):19–30. doi: 10.1042/bj1350019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Meaden P., Hill K., Wagner J., Slipetz D., Sommer S. S., Bussey H. The yeast KRE5 gene encodes a probable endoplasmic reticulum protein required for (1----6)-beta-D-glucan synthesis and normal cell growth. Mol Cell Biol. 1990 Jun;10(6):3013–3019. doi: 10.1128/mcb.10.6.3013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Nakayama K., Nagasu T., Shimma Y., Kuromitsu J., Jigami Y. OCH1 encodes a novel membrane bound mannosyltransferase: outer chain elongation of asparagine-linked oligosaccharides. EMBO J. 1992 Jul;11(7):2511–2519. doi: 10.1002/j.1460-2075.1992.tb05316.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Paravicini G., Cooper M., Friedli L., Smith D. J., Carpentier J. L., Klig L. S., Payton M. A. The osmotic integrity of the yeast cell requires a functional PKC1 gene product. Mol Cell Biol. 1992 Nov;12(11):4896–4905. doi: 10.1128/mcb.12.11.4896. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Raymond C. K., O'Hara P. J., Eichinger G., Rothman J. H., Stevens T. H. Molecular analysis of the yeast VPS3 gene and the role of its product in vacuolar protein sorting and vacuolar segregation during the cell cycle. J Cell Biol. 1990 Sep;111(3):877–892. doi: 10.1083/jcb.111.3.877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Raymond M., Gros P., Whiteway M., Thomas D. Y. Functional complementation of yeast ste6 by a mammalian multidrug resistance mdr gene. Science. 1992 Apr 10;256(5054):232–234. doi: 10.1126/science.1348873. [DOI] [PubMed] [Google Scholar]
  26. Roemer T., Bussey H. Yeast beta-glucan synthesis: KRE6 encodes a predicted type II membrane protein required for glucan synthesis in vivo and for glucan synthase activity in vitro. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11295–11299. doi: 10.1073/pnas.88.24.11295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Roemer T., Delaney S., Bussey H. SKN1 and KRE6 define a pair of functional homologs encoding putative membrane proteins involved in beta-glucan synthesis. Mol Cell Biol. 1993 Jul;13(7):4039–4048. doi: 10.1128/mcb.13.7.4039. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Roncero C., Valdivieso M. H., Ribas J. C., Durán A. Isolation and characterization of Saccharomyces cerevisiae mutants resistant to Calcofluor white. J Bacteriol. 1988 Apr;170(4):1950–1954. doi: 10.1128/jb.170.4.1950-1954.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rothstein R. J. One-step gene disruption in yeast. Methods Enzymol. 1983;101:202–211. doi: 10.1016/0076-6879(83)01015-0. [DOI] [PubMed] [Google Scholar]
  30. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Silverman S. J., Sburlati A., Slater M. L., Cabib E. Chitin synthase 2 is essential for septum formation and cell division in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1988 Jul;85(13):4735–4739. doi: 10.1073/pnas.85.13.4735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Valdivieso M. H., Mol P. C., Shaw J. A., Cabib E., Durán A. CAL1, a gene required for activity of chitin synthase 3 in Saccharomyces cerevisiae. J Cell Biol. 1991 Jul;114(1):101–109. doi: 10.1083/jcb.114.1.101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Van Rinsum J., Klis F. M., van den Ende H. Cell wall glucomannoproteins of Saccharomyces cerevisiae mnn9. Yeast. 1991 Oct;7(7):717–726. doi: 10.1002/yea.320070707. [DOI] [PubMed] [Google Scholar]
  34. Whiteway M., Szostak J. W. The ARD1 gene of yeast functions in the switch between the mitotic cell cycle and alternative developmental pathways. Cell. 1985 Dec;43(2 Pt 1):483–492. doi: 10.1016/0092-8674(85)90178-3. [DOI] [PubMed] [Google Scholar]
  35. von Heijne G. How signal sequences maintain cleavage specificity. J Mol Biol. 1984 Feb 25;173(2):243–251. doi: 10.1016/0022-2836(84)90192-x. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES