Publication:
Presence of a large β(1-3)glucan linked to chitin at the Saccharomyces cerevisiae mother-bud neck suggests involvement in localized growth control

Loading...
Thumbnail Image
Full text at PDC
Publication Date
2012-02-24
Advisors (or tutors)
Editors
Journal Title
Journal ISSN
Volume Title
Publisher
American Society for Microbiology
Citations
Google Scholar
Research Projects
Organizational Units
Journal Issue
Abstract
Previous results suggested that the chitin ring present at the yeast mother-bud neck, which is linked specifically to the nonreducing ends of β(1-3)glucan, may help to suppress cell wall growth at the neck by competing with β(1-6)glucan and thereby with mannoproteins for their attachment to the same sites. Here we explored whether the linkage of chitin to β(1-3)glucan may also prevent the remodeling of this polysaccharide that would be necessary for cell wall growth. By a novel mild procedure, β(1-3)glucan was isolated from cell walls, solubilized by carboxymethylation, and fractionated by size exclusion chromatography, giving rise to a very high-molecular-weight peak and to highly polydisperse material. The latter material, soluble in alkali, may correspond to glucan being remodeled, whereas the large-size fraction would be the final cross-linked structural product. In fact, the β(1-3)glucan of buds, where growth occurs, is solubilized by alkali. A gas1 mutant with an expected defect in glucan elongation showed a large increase in the polydisperse fraction. By a procedure involving sodium hydroxide treatment, carboxymethylation, fractionation by affinity chromatography on wheat germ agglutinin-agarose, and fractionation by size chromatography on Sephacryl columns, it was shown that the β(1-3)glucan attached to chitin consists mostly of high-molecular-weight material. Therefore, it appears that linkage to chitin results in a polysaccharide that cannot be further remodeled and does not contribute to growth at the neck. In the course of these experiments, the new finding was made that part of the chitin forms a noncovalent complex with β(1-3)glucan.
Description
Keywords
Citation
1. Bom IJ, et al. 1998. A new tool for studying the molecular architecture of the fungal cell wall: one step purification of recombinant Trichoderma _-(1-6)-glucanase expressed in Pichia pastoris. Biochim. Biophys. Acta 1425:419–424. 2. Cabib E. 2009. Two novel techniques for determination of polysaccharide cross-links show that Crh1p and Crh2p attach chitin to both _(1-6)- and _(1-3)glucan in the Saccharomyces cerevisiae cell wall. Eukaryot. Cell 8:1626 –1636. 3. Cabib E, Blanco N, Grau C, Rodríguez-Peña JM, Arroyo J. 2007. Crh1p and Crh2p are required for the cross-linking of chitin to _(1-6)glucan in the Saccharomyces cerevisiae cell wall. Mol. Microbiol. 63:921–935. 4. Cabib E, Durán A. 2005. Synthase III-dependent chitin is bound to different acceptors depending on location on the cell wall of budding yeast. J. Biol. Chem. 280:9170 –9179. 5. Cabib E, Roh D-H, Schmidt M, Crotti LB, Varma A. 2001. The yeast cell wall and septum as paradigms of cell growth and morphogenesis. J. Biol. Chem. 276:19679 –19682. 6. Caudron F, Barral Y. 2009. Septins and the lateral compartmentation of eukaryotic membranes. Dev. Cell 16:493–506. 7. Crotti LB, Drgon T, Cabib E. 2001. Yeast cell permeabilization by osmotic shock allows determination of enzymatic activities in situ. Anal. Biochem. 292:8 –16. 8. Drgonová J, Drgon T, Roh D-H, Cabib E. 1999. The GTP-binding protein Rho1p is required for cell cycle progression and polarization of the yeast cell. J. Cell Biol. 146:373–387. 9. Kapteyn JC, et al. 1997. Altered extent of cross-linking of _1,6-glucosylated mannoproteins to chitin in Saccharomyces cerevisiae mutants with reduced cell wall _1,3-glucan content. J. Bacteriol. 179:6279–6284. 10. Kitamura K, Kaneko T, Yamamoto Y. 1974. Lysis of viable yeast cells by enzymes of Arthrobacter luteus. II. Purification and properties of an enzyme, Zymolyase, which lyses viable yeast cells. J. Gen. Appl. Microbiol 20. 323–344. 11. Klis FM, Boorsma A, De Groot PWJ. 2006. Cell wall construction in Saccharomyces cerevisiae. Yeast 23:185–202. 12. Kollár R, Petráková E, Ashwell G, Robbins PW, Cabib E. 1995. Architecture of the yeast cell wall. The linkage between chitin and _(1¡3)-glucan. J. Biol. Chem. 270:1170 –1178. 13. Kollár R, et al. 1997. Architecture of the yeast cell wall. _(1¡6)-Glucan interconnects mannoprotein, _(1¡3)-glucan, and chitin. J. Biol. Chem. 272:17762–17775. 14. Kopecká M, Phaff HG, Fleet GH. 1974. Demonstration of a fibrillar component in the cell wall of the yeast Saccharomyces cerevisiae and its chemical nature. J. Cell Biol. 62:66 –76. 15. Lesage G, Bussey H. 2006. Cell wall assembly in Saccharomyces cerevisiae. Microbiol. Mol. Biol. Rev. 70:317–343. 16. Lippincott J, Li R. 1998. Sequential assembly of myosin II, an IQGAP-like protein, and filamentous actin to a ring structure involved in budding yeast cytokinesis. J. Cell Biol. 140:355–366. 17. Magnelli P, Cipollo JF, Abeijón C. 2002. A refined method for the determination of Saccharomyces cerevisiae cell wall composition and _-1,6-glucan fine structure. Anal. Biochem. 301:136 –150. 18. Manners DJ, Masson AJ, Patterson JC. 1973. The structure of a _-(1¡3)-D-glucan from yeast cell walls. Biochem. J. 135:19 –30. 19. Mol PC, Wessels JGH. 1987. Linkages between glucosaminoglycan and glucan determine alkali-insolubility of the glucan in walls of Saccharomyces cerevisiae. FEMS Microbiol. Lett. 41:95–99. 20. Mouyna I, et al. 2000. Glycosylphosphatidylinositol-anchored glucanosyltransferases play an active role in the biosynthesis of the fungal cell wall. J. Biol. Chem. 275:14882–14889. 21. Popolo L, Gilardelli D, Bonfante P, Vai M. 1997. Increase in chitin as an essential response to defects in assembly of cell wall polymers in the ggp1_mutant of Saccharomyces cerevisiae. J. Bacteriol. 179:463– 469. 22. Roberts RL, Cabib E. 1982. Serratia marcescens chitinase: one-step purification and use for the determination of chitin. Anal. Biochem. 127:402–412. 23. Rodríguez-Peña JM, Cid VJ, Arroyo J, Nombela C. 2000. A novel family of cell wall-related proteins regulated differentially during the yeast life cycle. Mol. Cell. Biol. 20:3245–3255. 24. Rodríguez-Peña JM, Rodríguez C, Alvarez A, Nombela C, Arroyo J. 2002. Mechanisms for targeting of the Saccharomyces cerevisiae GPIanchored cell wall protein Crh2p to polarized growth sites. J. Cell Sci. 115:2549 –2558. 25. Rolli E, et al. 2009. Immobilization of the glycosylphosphatidylinositolanchored Gas1 protein into the chitin ring and septum is required for proper morphogenesis in yeast. Mol. Biol. Cell 20:4856–4870. 26. Schmidt M, Bowers B, Varma A, Roh D-H, Cabib E. 2002. In budding yeast, contraction of the actomyosin ring and formation of the primary septum at cytokinesis depend on each other. J. Cell Sci. 115:293–302. 27. Schmidt M, Varma A, Drgon T, Bowers B, Cabib E. 2003. Septins, under Cla4p regulation, and the chitin ring are required for neck integrity in budding yeast. Mol. Biol. Cell 14:2128 –2141. 28. Shaw JA, et al. 1991. The function of chitin synthases 2 and 3 in the Saccharomyces cerevisiae cell cycle. J. Cell Biol. 114:111–123. 29. Sikorski R, Hieter P. 1989. A system of shuttle vectors and host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics 122:19 –28. 30. Trevelyan WE, Harrison JS. 1952. Studies on yeast metabolism 1. Fractionation and microdetermination of cell carbohydrates. Biochem. J. 50: 298–303.
Collections