Skip to main content
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1983 May;3(5):922–930. doi: 10.1128/mcb.3.5.922

Chitin synthesis and localization in cell division cycle mutants of Saccharomyces cerevisiae.

R L Roberts, B Bowers, M L Slater, E Cabib
PMCID: PMC368615  PMID: 6223209

Abstract

Growth of Saccharomyces cerevisiae cell cycle mutants cdc3, cdc4, cdc7, cdc24, and cdc28 at a nonpermissive temperature (37 degrees C) resulted in increased accumulation of chitin relative to other cell wall components, as compared with that observed at a permissive temperature (25 degrees C). Wild-type cells showed the same chitin/carbohydrate ratio at both temperatures, whereas mutants cdc13 and cdc21 yielded only a small increase in the ratio at 37 degrees C. These results confirm and extend those reported by B. F. Sloat and J. R. Pringle (Science 200:1171-1173, 1978) for mutant cdc24. The distribution of chitin in the cell wall was studied by electron microscopy, by specific staining with wheat germ agglutinin-colloidal gold complexes. At the permissive temperature, chitin was restricted to the septal region in all strains, whereas at 37 degrees C a generalized distribution of chitin in the cell wall was observed in all mutants. These results do not support a unique interdependence between the product of the cdc24 gene and localization of chitin deposition; they suggest that unbalanced conditions created in the cell by arresting the cycle at different stages result in generalized activation of the chitin synthetase zymogen. Thus, blockage of an event in the cell cycle may lead to consequences that are not functionally related to that event under normal conditions.

Full text

PDF
922

Images in this article

Selected References

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

  1. Cabib E., Bowers B. Timing and function of chitin synthesis in yeast. J Bacteriol. 1975 Dec;124(3):1586–1593. doi: 10.1128/jb.124.3.1586-1593.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Culotti J., Hartwell L. H. Genetic control of the cell division cycle in yeast. 3. Seven genes controlling nuclear division. Exp Cell Res. 1971 Aug;67(2):389–401. doi: 10.1016/0014-4827(71)90424-1. [DOI] [PubMed] [Google Scholar]
  3. Duran A., Cabib E., Bowers B. Chitin synthetase distribution on the yeast plasma membrane. Science. 1979 Jan 26;203(4378):363–365. doi: 10.1126/science.366747. [DOI] [PubMed] [Google Scholar]
  4. Duran A., Cabib E. Solubilization and partial purification of yeast chitin synthetase. Confirmation of the zymogenic nature of the enzyme. J Biol Chem. 1978 Jun 25;253(12):4419–4425. [PubMed] [Google Scholar]
  5. Geoghegan W. D., Ackerman G. A. Adsorption of horseradish peroxidase, ovomucoid and anti-immunoglobulin to colloidal gold for the indirect detection of concanavalin A, wheat germ agglutinin and goat anti-human immunoglobulin G on cell surfaces at the electron microscopic level: a new method, theory and application. J Histochem Cytochem. 1977 Nov;25(11):1187–1200. doi: 10.1177/25.11.21217. [DOI] [PubMed] [Google Scholar]
  6. Hartwell L. H. Genetic control of the cell division cycle in yeast. II. Genes controlling DNA replication and its initiation. J Mol Biol. 1971 Jul 14;59(1):183–194. doi: 10.1016/0022-2836(71)90420-7. [DOI] [PubMed] [Google Scholar]
  7. Hartwell L. H. Genetic control of the cell division cycle in yeast. IV. Genes controlling bud emergence and cytokinesis. Exp Cell Res. 1971 Dec;69(2):265–276. doi: 10.1016/0014-4827(71)90223-0. [DOI] [PubMed] [Google Scholar]
  8. Hartwell L. H. Macromolecule synthesis in temperature-sensitive mutants of yeast. J Bacteriol. 1967 May;93(5):1662–1670. doi: 10.1128/jb.93.5.1662-1670.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hartwell L. H., Mortimer R. K., Culotti J., Culotti M. Genetic Control of the Cell Division Cycle in Yeast: V. Genetic Analysis of cdc Mutants. Genetics. 1973 Jun;74(2):267–286. doi: 10.1093/genetics/74.2.267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hartwell L. H. Three additional genes required for deoxyribonucleic acid synthesis in Saccharomyces cerevisiae. J Bacteriol. 1973 Sep;115(3):966–974. doi: 10.1128/jb.115.3.966-974.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Horisberger M., Vonlanthen M. Location of mannan and chitin on thin sections of budding yeasts with gold markers. Arch Microbiol. 1977 Oct 24;115(1):1–7. doi: 10.1007/BF00427837. [DOI] [PubMed] [Google Scholar]
  12. Minke R., Blackwell J. The structure of alpha-chitin. J Mol Biol. 1978 Apr 5;120(2):167–181. doi: 10.1016/0022-2836(78)90063-3. [DOI] [PubMed] [Google Scholar]
  13. Molano J., Bowers B., Cabib E. Distribution of chitin in the yeast cell wall. An ultrastructural and chemical study. J Cell Biol. 1980 May;85(2):199–212. doi: 10.1083/jcb.85.2.199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Molano J., Durán A., Cabib E. A rapid and sensitive assay for chitinase using tritiated chitin. Anal Biochem. 1977 Dec;83(2):648–656. doi: 10.1016/0003-2697(77)90069-0. [DOI] [PubMed] [Google Scholar]
  15. REISSIG J. L., STORMINGER J. L., LELOIR L. F. A modified colorimetric method for the estimation of N-acetylamino sugars. J Biol Chem. 1955 Dec;217(2):959–966. [PubMed] [Google Scholar]
  16. REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Schekman R., Brawley V. Localized deposition of chitin on the yeast cell surface in response to mating pheromone. Proc Natl Acad Sci U S A. 1979 Feb;76(2):645–649. doi: 10.1073/pnas.76.2.645. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sloat B. F., Adams A., Pringle J. R. Roles of the CDC24 gene product in cellular morphogenesis during the Saccharomyces cerevisiae cell cycle. J Cell Biol. 1981 Jun;89(3):395–405. doi: 10.1083/jcb.89.3.395. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sloat B. F., Pringle J. R. A mutant of yeast defective in cellular morphogenesis. Science. 1978 Jun 9;200(4346):1171–1173. doi: 10.1126/science.349694. [DOI] [PubMed] [Google Scholar]
  20. TREVELYAN W. E., HARRISON J. S. Studies on yeast metabolism. I. Fractionation and microdetermination of cell carbohydrates. Biochem J. 1952 Jan;50(3):298–303. doi: 10.1042/bj0500298. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ulane R. E., Cabib E. The activating system of chitin synthetase from Saccharomyces cerevisiae. Purification and properties of the activating factor. J Biol Chem. 1976 Jun 10;251(11):3367–3374. [PubMed] [Google Scholar]
  22. Wolf D. H., Ehmann C. Studies on a proteinase B mutant of yeast. Eur J Biochem. 1979 Aug 1;98(2):375–384. doi: 10.1111/j.1432-1033.1979.tb13197.x. [DOI] [PubMed] [Google Scholar]
  23. Zubenko G. S., Mitchell A. P., Jones E. W. Mapping of the proteinase b structural gene PRB1, in Saccharomyces cerevisiae and identification of nonsense alleles within the locus. Genetics. 1980 Sep;96(1):137–146. doi: 10.1093/genetics/96.1.137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Zubenko G. S., Mitchell A. P., Jones E. W. Septum formation, cell division, and sporulation in mutants of yeast deficient in proteinase B. Proc Natl Acad Sci U S A. 1979 May;76(5):2395–2399. doi: 10.1073/pnas.76.5.2395. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES