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Proc Natl Acad Sci U S A. Dec 15, 1991; 88(24): 11295–11299.
PMCID: PMC53121

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.

Abstract

The KRE6 gene product is required for synthesis of the major beta-glucans of the yeast cell wall, as mutations in this gene confer reduced levels of both the (1----6)- and (1----3)-beta-D-glucan polymers. Cloning and sequencing of KRE6 reveals a gene encoding a predicted 80-kDa protein with a central transmembrane domain and the topology of a type II membrane protein. Null mutants of KRE6 grow slowly, have larger cells, and show a reduction in alkali-insoluble wall glucans. The mutants show good viability and are not osmotically sensitive, but they are more susceptible to beta-glucanase digestion and mechanical stress than wild-type cells. The specific activity of the GTP-dependent, membrane-associated, in vitro (1----3)-beta-glucan synthase is reduced 50% in kre6 null mutants, and this reduction correlates with the mutation in meiotic tetrads. Transformants of kre6 null mutants with a KRE6 gene expressed from a centomere-based vector show a 4- to 5-fold increase in in vitro (1----3)-beta-glucan synthase activity over transformants with the vector alone. The phenotype and structure of the KRE6 product, Kre6p, suggest that Kre6p may be a beta-glucan synthase, and if so, it implies that beta-glucan synthases are functionally redundant in yeast. Alternatively, Kre6p may be part of a single multiprotein glucan synthase or modulate its activity. Use of KRE6 should permit a genetic analysis of eukaryotic (1----3)-beta-glucan synthesis.

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  • Manners DJ, Masson AJ, Patterson JC. The structure of a beta-(1 leads to 3)-D-glucan from yeast cell walls. Biochem J. 1973 Sep;135(1):19–30. [PMC free article] [PubMed]
  • Manners DJ, Masson AJ, Patterson JC, 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. [PMC free article] [PubMed]
  • Wong HC, Fear AL, Calhoon RD, Eichinger GH, Mayer R, Amikam D, Benziman M, Gelfand DH, Meade JH, Emerick AW, et al. Genetic organization of the cellulose synthase operon in Acetobacter xylinum. Proc Natl Acad Sci U S A. 1990 Oct;87(20):8130–8134. [PMC free article] [PubMed]
  • Saxena IM, Lin FC, Brown RM., Jr Cloning and sequencing of the cellulose synthase catalytic subunit gene of Acetobacter xylinum. Plant Mol Biol. 1990 Nov;15(5):673–683. [PubMed]
  • Frost DJ, Read SM, Drake RR, Haley BE, Wasserman BP. Identification of the UDP-glucose-binding polypeptide of callose synthase from Beta vulgaris L. by photoaffinity labeling with 5-azido-UDP-glucose. J Biol Chem. 1990 Feb 5;265(4):2162–2167. [PubMed]
  • Kang MS, Cabib E. Regulation of fungal cell wall growth: a guanine nucleotide-binding, proteinaceous component required for activity of (1----3)-beta-D-glucan synthase. Proc Natl Acad Sci U S A. 1986 Aug;83(16):5808–5812. [PMC free article] [PubMed]
  • Huffaker TC, Robbins PW. Yeast mutants deficient in protein glycosylation. Proc Natl Acad Sci U S A. 1983 Dec;80(24):7466–7470. [PMC free article] [PubMed]
  • Bulawa CE, Slater M, Cabib E, Au-Young J, Sburlati A, Adair WL, Jr, Robbins PW. The S. cerevisiae structural gene for chitin synthase is not required for chitin synthesis in vivo. Cell. 1986 Jul 18;46(2):213–225. [PubMed]
  • Silverman SJ, Sburlati A, Slater ML, 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. [PMC free article] [PubMed]
  • Bulawa CE, Osmond BC. Chitin synthase I and chitin synthase II are not required for chitin synthesis in vivo in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7424–7428. [PMC free article] [PubMed]
  • Farkas I, Hardy TA, DePaoli-Roach AA, Roach PJ. Isolation of the GSY1 gene encoding yeast glycogen synthase and evidence for the existence of a second gene. J Biol Chem. 1990 Dec 5;265(34):20879–20886. [PubMed]
  • Meaden P, Hill K, Wagner J, Slipetz D, Sommer SS, 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. [PMC free article] [PubMed]
  • Murray AW, Szostak JW. Pedigree analysis of plasmid segregation in yeast. Cell. 1983 Oct;34(3):961–970. [PubMed]
  • Nakajima T, Ballou CE. Characterization of the carbohydrate fragments obtained from Saccharomyces cerevisiae mannan by alkaline degradation. J Biol Chem. 1974 Dec 10;249(23):7679–7684. [PubMed]
  • 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]
  • 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. [PMC free article] [PubMed]
  • 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. [PMC free article] [PubMed]
  • Sikorski RS, Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. [PMC free article] [PubMed]
  • Rose MD, Novick P, Thomas JH, Botstein D, Fink GR. A Saccharomyces cerevisiae genomic plasmid bank based on a centromere-containing shuttle vector. Gene. 1987;60(2-3):237–243. [PubMed]
  • Vernet T, Dignard D, Thomas DY. A family of yeast expression vectors containing the phage f1 intergenic region. Gene. 1987;52(2-3):225–233. [PubMed]
  • Hoffman CS, Winston F. A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. Gene. 1987;57(2-3):267–272. [PubMed]
  • Huisman O, Raymond W, Froehlich KU, Errada P, Kleckner N, Botstein D, Hoyt MA. A Tn10-lacZ-kanR-URA3 gene fusion transposon for insertion mutagenesis and fusion analysis of yeast and bacterial genes. Genetics. 1987 Jun;116(2):191–199. [PMC free article] [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PMC free article] [PubMed]
  • Rothstein RJ. One-step gene disruption in yeast. Methods Enzymol. 1983;101:202–211. [PubMed]
  • Boone C, Sommer SS, 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. [PMC free article] [PubMed]
  • Cabib E, Kang MS. Fungal 1,3-beta-glucan synthase. Methods Enzymol. 1987;138:637–642. [PubMed]
  • Szaniszlo PJ, Kang MS, Cabib E. Stimulation of beta(1----3)glucan synthetase of various fungi by nucleoside triphosphates: generalized regulatory mechanism for cell wall biosynthesis. J Bacteriol. 1985 Mar;161(3):1188–1194. [PMC free article] [PubMed]
  • Shematek EM, Braatz JA, Cabib E. Biosynthesis of the yeast cell wall. I. Preparation and properties of beta-(1 leads to 3)glucan synthetase. J Biol Chem. 1980 Feb 10;255(3):888–894. [PubMed]
  • Wickner WT, Lodish HF. Multiple mechanisms of protein insertion into and across membranes. Science. 1985 Oct 25;230(4724):400–407. [PubMed]
  • Parks GD, Lamb RA. Topology of eukaryotic type II membrane proteins: importance of N-terminal positively charged residues flanking the hydrophobic domain. Cell. 1991 Feb 22;64(4):777–787. [PubMed]
  • Paulson JC, Colley KJ. Glycosyltransferases. Structure, localization, and control of cell type-specific glycosylation. J Biol Chem. 1989 Oct 25;264(30):17615–17618. [PubMed]

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