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Changes in cell wall composition of deformedras1 cells ofSchizosaccharomyces pombe

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Abstract

Disruption of theSchizosaccharomyces pombe ras1 gene results in a morphological transformation to large spheres, in contrast to wild-type cells which grow as rods. Chemical analysis of isolated cell walls showed no significant changes in saccharide content but an increase in protein and phosphate contents inras1 walls relative to parent walls. Polymers tightly bound to the cell wall were solubilized by SDS treatment. Several compounds with molar mass ranging from 22 to 130 kDa and more were resolved by gel filtration and SDS-PAGE. Among low-molar-mass species, a component moving as a band at 31 kDa was conspicuous inras1 cell walls. It was solubilized by heating in Tris-HCl buffer and shown to have a β-1,3-glucanase activity against laminarin. The level of the enzyme was by 30% higher in theras1 cell wall than in the wild-type cell wall. This enzyme may participate in the remodelling of the rigid glucan network and account (at least partially) for the aberrant cell shape. Theras1 cell wall contained a high level of charged polymers, especially phosphoproteins, raising the appealing possibility thatras1 is involved in a putative kinase cascade required to sense and respond to external stimuli destined for the cell wall. Although the present study shows thatras1 loss of function and altered cell wall composition are closely linked defects, it has still to be shown that theras1 protein is directly involved in alterations found in the mutant cell walls.

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References

  • Bacon J.S.D., Jones D., Farmer V.C., Webley D.M.: The occurrence of α-(1→3) glucan inCryptococcus, Schizosaccharomyces andPolyporus species and its hydrolysis by a streptomycete culture filtrate lysing cell walls ofCryptococcus.Biochim. Biophys. Acta158, 313–315 (1968).

    PubMed  CAS  Google Scholar 

  • Bush D.A., Horisberger M., Horman I., Wursch P.: The cell wall structure ofSchizosaccharomyces pombe.J. Gen. Microbiol.81, 199–206 (1974).

    PubMed  CAS  Google Scholar 

  • Cawley T.N., Harrington M.G., Letters R.: A study of the phosphate linkages in phosphomannan in cell wall ofSaccharomyces cerevisiæ.Biochem. J.129, 711–720 (1972).

    PubMed  CAS  Google Scholar 

  • Dubois A., Gilles K.A., Hamilton K.J.: Coloric method for determination of sugars and related substances.Anal. Chem.28, 350–356 (1972).

    Article  Google Scholar 

  • Errede B., Levin D.E.: A conserved kinase cascade for MAP kinase activation ion yeast.Curr. Opinion Cell Biol.5, 254–260 (1993).

    Article  PubMed  CAS  Google Scholar 

  • Fink G.R.: The biochemical genetics of yeast.Methods Enzymol.17, 59–78 (1970).

    Article  CAS  Google Scholar 

  • Font de Mora J., Valentin E., Herrero E., Sentandreu R.: Glycoprotein molecules in the cell wall ofSchizosaccharomyces pombe wild-type cells and a morphologically altered mutant resistant to papulacandin B.J. Gen. Microbiol.136, 2251–2259 (1990).

    PubMed  CAS  Google Scholar 

  • Fukui Y., Kaziro Y.: Molecular cloning and sequence analysis of aras gene fromSchizosaccharomyces pombe.EMBO J.4, 687–691 (1985).

    PubMed  CAS  Google Scholar 

  • Ghuysen J.M., Tipper D.J., Birge C.H., Strominger J.L.: Structure of the cell wall ofStaphylococcus aureus, strain Copenhagen. The soluble glycopeptide and its sequential degradation by peptidase.Biochemistry4, 2245–2254 (1965).

    Article  CAS  Google Scholar 

  • Ghuysen J.M., Tipper D.J., Strominger J.L.: Enzymes that degrade bacterial cell wall.Methods Enzymol.8, 685–699 (1966).

    CAS  Google Scholar 

  • Herrero E., Sanz P., Sentandreu R.: Cell wall proteins liberated by Zymolase from several ascomycetous and imperfect yeasts.J. Gen. Microbiol.133, 2895–2903 (1987).

    CAS  Google Scholar 

  • Horisberger M., Rosset J.: Localization of ì-galactomannan on the surface ofSchizosaccharomyces pombe cells by scanning electron microscopy.Arch. Microbiol.112, 123–126 (1977).

    Article  CAS  Google Scholar 

  • Horisberger M., Volanthen M., Rosset J.: Localization of ì-galactomannan and of wheat agglutinin receptors inSchizosaccharomyces pombe.Arch. Microbiol.119, 107–111 (1978).

    Article  PubMed  CAS  Google Scholar 

  • Hughes D.A., Imai Y., Yamamoto M.: Regulation of theras pathway in the fission yeastSchizosaccharomyces pombe, pp. 41–47 in D.A. Spandidos (Ed.):The Superfamily of rasRelated Genes. Plenum Press, New York 1991.

    Google Scholar 

  • Klebl F., Tanner W.: Molecular cloning of cell wall exo-β-1,3-glucanase fromSaccharomyces cerevisiœ.J. Bacteriol.171, 6259–6264 (1989).

    PubMed  CAS  Google Scholar 

  • Kobori H., Toda T., Yaguchi H., Toya M., Yanagida M., Osumi M.: Fission yeast protein kinase C gene homologues are required for protoplast regeneration: a functional link between cell wall formation and cell shape control.J. Cell Sci.107, 1131–1136 (1994).

    PubMed  CAS  Google Scholar 

  • Laemmli U.K.: Cleavage of structural proteins during assembly of the head of bacteriophage T4.Nature (London)227, 680–685 (1970).

    Article  CAS  Google Scholar 

  • Manners D.J., Meyer M.T.: The molecular structure of some glucans from the cell wall ofSchizosaccharomyces pombe.Carbohydr. Res.57, 189–203 (1977).

    Article  CAS  Google Scholar 

  • Manners D.J., Masson A.J., Patterson J.C.: The heterogeneity of glucan preparations from the walls of various yeasts.J. Gen. Microbiol.80, 411–417 (1974).

    PubMed  CAS  Google Scholar 

  • McClare C.W.F.: An accurate and convenient organic phosphorus assay.Anal. Biochem.39, 527–530 (1971).

    Article  PubMed  CAS  Google Scholar 

  • Mendoza G., Villanueva J.R.: Preparation of cell walls of yeast.Can. J. Microbiol.9, 141–144 (1963).

    Article  Google Scholar 

  • Mrša B., Klebl F., Tanner W.: Purification and characterization of theSaccharomyces cerevisiœ BGL2 gene product, a cell wall endo-β-1,3-glucanase.J. Bacteriol.175, 2102–2106 (1993).

    PubMed  Google Scholar 

  • Neiman A.M.: Conservation and reiteration of a kinase cascade.Trends Genet.9, 390–394 (1993).

    Article  PubMed  CAS  Google Scholar 

  • Nielsen O.: Signal transduction during mating and meiosis inS. pombe.Trends Cell Biol.3, 60–65 (1993).

    Article  PubMed  CAS  Google Scholar 

  • Park J.T., Johnson M.J.: A submicrodetermination of glucose.J. Biol. Chem.181, 149–151 (1949).

    PubMed  CAS  Google Scholar 

  • Pastor F.I.J., Valentin E., Herrero E., Sentandreu R.: Structure of theSaccharomyces cerevisiœ cell wall architecture.Biochim. Biophys. Acta802, 292–300 (1984).

    CAS  Google Scholar 

  • Pichová A., Streiblová E.: Features of the periphery in the deformedras1 cells ofSchizosaccharomyces pombe.Exp. Mycol.16, 178–187 (1992).

    Article  Google Scholar 

  • Pichová A., Vondráková D., Streiblová E.: Reversion of deformedras1 cells ofSchizosaccharomyces pombe to rod-like cells by nutrient stimulation.Acta Bot. Neerl.43, 65–69 (1994).

    Google Scholar 

  • Ribas J.C., Roncero C., Rico H., Duran A.: Characterization of aSchizosaccharomyces pombe morphological mutant altered in the galactomannan content.FEMS Microbiol. Lett.97, 263–268 (1991).

    Article  Google Scholar 

  • Sawardeker J.S., Sloneker J.H., Jeanes A.: Quantitative determination of monosaccharides and their alditol acetates by gas chromatography.Anal. Chem.57, 1602–1604 (1965).

    Article  Google Scholar 

  • Sietsma J.H., Wessels J.G.H.: The occurrence of glucosaminoglycan in the wall ofSchizosaccharomyces pombe.J. Gen. Microbiol.136, 2261–2265 (1990).

    PubMed  CAS  Google Scholar 

  • Streiblová E.: Fission, pp. 79–92 in J.W. Arnold (Ed.):Yeast Envelopes: Biochemistry, Biophysics and Ultrastructure. CRC Press, Boca Raton (Florida) 1981.

    Google Scholar 

  • Toda T., Shimanuki M., Yanagida M.: Two novel protein kinase C related genes of fission yeast are essential for cell viability and implicated in cell shape control.EMBO J.12, 1987–1995 (1993).

    PubMed  CAS  Google Scholar 

  • Valentin E., Herrero E., Pastor F.I.J., Sentandreu R.: Solubilization and analysis of mannoprotein molecules from the cell wall ofSaccharomyces cerevisiœ.J. Gen. Microbiol.130, 1419–1428 (1984).

    CAS  Google Scholar 

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Harmouch, N., Pichová, A., Coulon, J. et al. Changes in cell wall composition of deformedras1 cells ofSchizosaccharomyces pombe . Folia Microbiol 40, 519–527 (1995). https://doi.org/10.1007/BF02814736

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