Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1994 Dec;14(12):7685–7694. doi: 10.1128/mcb.14.12.7685

Are yeast chitin synthases regulated at the transcriptional or the posttranslational level?

W J Choi 1, B Santos 1, A Durán 1, E Cabib 1
PMCID: PMC359310  PMID: 7969112

Abstract

The three chitin synthases of Saccharomyces cerevisiae, Chs1, Chs2, and Chs3, participate in septum and cell wall formation of vegetative cells and in wall morphogenesis of conjugating cells and spores. Because of the differences in the nature and in the time of execution of their functions, the synthases must be specifically and individually regulated. The nature of that regulation has been investigated by measuring changes in the levels of the three synthases and of the messages of the three corresponding genes, CHS1, CHS2, and CAL1/CSD2/DIT101/KTI2 (referred to below as CAL1/CSD2), during the budding and sexual cycles. By transferring cells carrying CHS2 under the control of a GAL1 promoter from galactose-containing medium to glucose-containing medium, transcription of CHS2 was shut off. This resulted in a rapid disappearance of Chs2, whereas the mRNA decayed much more slowly. Furthermore, Chs2 levels experienced pronounced oscillations during the budding cycle and were decreased in the sexual cycle, indicating that this enzyme is largely regulated by a process of synthesis and degradation. For CHS1 and CAL1/CSD2, however, a stop in transcription was followed by a slow decrease in the level of zymogen (Chs1) or an increase in the level of activity (Chs3), despite a rapid drop in message level in both cases. In synchronized cultures, Chs1 levels were constant during the cell cycle. Thus, for Chs1 and Chs3, posttranslational regulation, probably by activation of latent forms, appears to be predominant. Since Chs2, like Chs1, is found in the cell in the zymogenic form, a posttranslational activation step appears to be necessary for this synthase also.

Full text

PDF
7685

Images in this article

7688Image
on p.7688
7689Image
on p.7689
7689Image
on p.7689
7691Image
on p.7691
7691Image
on p.7691

Selected References

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

  1. Appeltauer U., Achstetter T. Hormone-induced expression of the CHS1 gene from Saccharomyces cerevisiae. Eur J Biochem. 1989 Apr 15;181(1):243–247. doi: 10.1111/j.1432-1033.1989.tb14718.x. [DOI] [PubMed] [Google Scholar]
  2. Briza P., Ellinger A., Winkler G., Breitenbach M. Chemical composition of the yeast ascospore wall. The second outer layer consists of chitosan. J Biol Chem. 1988 Aug 15;263(23):11569–11574. [PubMed] [Google Scholar]
  3. 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]
  4. Bulawa C. E. Genetics and molecular biology of chitin synthesis in fungi. Annu Rev Microbiol. 1993;47:505–534. doi: 10.1146/annurev.mi.47.100193.002445. [DOI] [PubMed] [Google Scholar]
  5. Bulawa C. E., Osmond B. C. 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. doi: 10.1073/pnas.87.19.7424. [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. Cabib E. Differential inhibition of chitin synthetases 1 and 2 from Saccharomyces cerevisiae by polyoxin D and nikkomycins. Antimicrob Agents Chemother. 1991 Jan;35(1):170–173. doi: 10.1128/aac.35.1.170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cabib E., Sburlati A., Bowers B., Silverman S. J. Chitin synthase 1, an auxiliary enzyme for chitin synthesis in Saccharomyces cerevisiae. J Cell Biol. 1989 May;108(5):1665–1672. doi: 10.1083/jcb.108.5.1665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cabib E., Silverman S. J., Shaw J. A. Chitinase and chitin synthase 1: counterbalancing activities in cell separation of Saccharomyces cerevisiae. J Gen Microbiol. 1992 Jan;138(1):97–102. doi: 10.1099/00221287-138-1-97. [DOI] [PubMed] [Google Scholar]
  10. Carmichael G. G., McMaster G. K. The analysis of nucleic acids in gels using glyoxal and acridine orange. Methods Enzymol. 1980;65(1):380–391. doi: 10.1016/s0076-6879(80)65049-6. [DOI] [PubMed] [Google Scholar]
  11. Choi W. J., Cabib E. The use of divalent cations and pH for the determination of specific yeast chitin synthetases. Anal Biochem. 1994 Jun;219(2):368–372. doi: 10.1006/abio.1994.1278. [DOI] [PubMed] [Google Scholar]
  12. Choi W. J., Sburlati A., Cabib E. Chitin synthase 3 from yeast has zymogenic properties that depend on both the CAL1 and the CAL3 genes. Proc Natl Acad Sci U S A. 1994 May 24;91(11):4727–4730. doi: 10.1073/pnas.91.11.4727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Correa J. U., Elango N., Polacheck I., Cabib E. Endochitinase, a mannan-associated enzyme from Saccharomyces cerevisiae. J Biol Chem. 1982 Feb 10;257(3):1392–1397. [PubMed] [Google Scholar]
  14. Domdey H., Apostol B., Lin R. J., Newman A., Brody E., Abelson J. Lariat structures are in vivo intermediates in yeast pre-mRNA splicing. Cell. 1984 Dec;39(3 Pt 2):611–621. doi: 10.1016/0092-8674(84)90468-9. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Finkelstein D. B., Strausberg S. Metabolism of alpha-factor by a mating type cells of Saccharomyces cerevisiae. J Biol Chem. 1979 Feb 10;254(3):796–803. [PubMed] [Google Scholar]
  17. 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]
  18. Kang M. S., Elango N., Mattia E., Au-Young J., Robbins P. W., Cabib E. Isolation of chitin synthetase from Saccharomyces cerevisiae. Purification of an enzyme by entrapment in the reaction product. J Biol Chem. 1984 Dec 10;259(23):14966–14972. [PubMed] [Google Scholar]
  19. Kronstad J. W., Holly J. A., MacKay V. L. A yeast operator overlaps an upstream activation site. Cell. 1987 Jul 31;50(3):369–377. doi: 10.1016/0092-8674(87)90491-0. [DOI] [PubMed] [Google Scholar]
  20. Kuranda M. J., Robbins P. W. Chitinase is required for cell separation during growth of Saccharomyces cerevisiae. J Biol Chem. 1991 Oct 15;266(29):19758–19767. [PubMed] [Google Scholar]
  21. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  22. Ng R., Abelson J. Isolation and sequence of the gene for actin in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3912–3916. doi: 10.1073/pnas.77.7.3912. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nurse P. Universal control mechanism regulating onset of M-phase. Nature. 1990 Apr 5;344(6266):503–508. doi: 10.1038/344503a0. [DOI] [PubMed] [Google Scholar]
  24. Ohya Y., Goebl M., Goodman L. E., Petersen-Bjørn S., Friesen J. D., Tamanoi F., Anraku Y. Yeast CAL1 is a structural and functional homologue to the DPR1 (RAM) gene involved in ras processing. J Biol Chem. 1991 Jul 5;266(19):12356–12360. [PubMed] [Google Scholar]
  25. Ohya Y., Ohsumi Y., Anraku Y. Genetic study of the role of calcium ions in the cell division cycle of Saccharomyces cerevisiae: a calcium-dependent mutant and its trifluoperazine-dependent pseudorevertants. Mol Gen Genet. 1984;193(3):389–394. doi: 10.1007/BF00382073. [DOI] [PubMed] [Google Scholar]
  26. Orlean P. Two chitin synthases in Saccharomyces cerevisiae. J Biol Chem. 1987 Apr 25;262(12):5732–5739. [PubMed] [Google Scholar]
  27. Pammer M., Briza P., Ellinger A., Schuster T., Stucka R., Feldmann H., Breitenbach M. DIT101 (CSD2, CAL1), a cell cycle-regulated yeast gene required for synthesis of chitin in cell walls and chitosan in spore walls. Yeast. 1992 Dec;8(12):1089–1099. doi: 10.1002/yea.320081211. [DOI] [PubMed] [Google Scholar]
  28. Percival-Smith A., Segall J. Isolation of DNA sequences preferentially expressed during sporulation in Saccharomyces cerevisiae. Mol Cell Biol. 1984 Jan;4(1):142–150. doi: 10.1128/mcb.4.1.142. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Sburlati A., Cabib E. Chitin synthetase 2, a presumptive participant in septum formation in Saccharomyces cerevisiae. J Biol Chem. 1986 Nov 15;261(32):15147–15152. [PubMed] [Google Scholar]
  31. 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]
  32. Shaw J. A., Mol P. C., Bowers B., Silverman S. J., Valdivieso M. H., Durán A., Cabib E. The function of chitin synthases 2 and 3 in the Saccharomyces cerevisiae cell cycle. J Cell Biol. 1991 Jul;114(1):111–123. doi: 10.1083/jcb.114.1.111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. Trueheart J., Boeke J. D., Fink G. R. Two genes required for cell fusion during yeast conjugation: evidence for a pheromone-induced surface protein. Mol Cell Biol. 1987 Jul;7(7):2316–2328. doi: 10.1128/mcb.7.7.2316. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]

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

RESOURCES