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.
MeSH Terms
Aminoglycosides
Anti-Bacterial Agents/pharmacology
Cell Cycle
Cell Wall/metabolism
Chitin Synthase/genetics,metabolism
Gene Expression Regulation, Fungal
Protein Biosynthesis
RNA, Fungal/metabolism
RNA, Messenger/metabolism
Saccharomyces cerevisiae/enzymology
Spores, Fungal
Transcription, Genetic
Chemicals
Aminoglycosides
Anti-Bacterial Agents
RNA, Fungal
RNA, Messenger
nikkomycin
Chitin Synthase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Choi W J
Laboratory of Biochemistry and Metabolism, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, Maryland 20892.
Santos B
Durán A
Cabib E
References (35)
35 references, click to expand
-
The function of chitin synthases 2 and 3 in the Saccharomyces cerevisiae cell cycle.
J Cell Biol. 1991 Jul;114(1):111-23
PMID: 2050738
-
Genetics and molecular biology of chitin synthesis in fungi.
Annu Rev Microbiol. 1993;47:505-34
PMID: 8257107
-
Two genes required for cell fusion during yeast conjugation: evidence for a pheromone-induced surface protein.
Mol Cell Biol. 1987 Jul;7(7):2316-28
PMID: 3302672
-
Chitin synthase 1, an auxiliary enzyme for chitin synthesis in Saccharomyces cerevisiae.
J Cell Biol. 1989 May;108(5):1665-72
PMID: 2523889
-
Chitinase and chitin synthase 1: counterbalancing activities in cell separation of Saccharomyces cerevisiae.
J Gen Microbiol. 1992 Jan;138(1):97-102
PMID: 1556560
-
Isolation and sequence of the gene for actin in Saccharomyces cerevisiae.
Proc Natl Acad Sci U S A. 1980 Jul;77(7):3912-6
PMID: 7001447
-
Isolation of DNA sequences preferentially expressed during sporulation in Saccharomyces cerevisiae.
Mol Cell Biol. 1984 Jan;4(1):142-50
PMID: 6366508
-
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):4735-9
PMID: 2968606
-
The analysis of nucleic acids in gels using glyoxal and acridine orange.
Methods Enzymol. 1980;65(1):380-91
PMID: 6154866
-
Metabolism of alpha-factor by a mating type cells of Saccharomyces cerevisiae.
J Biol Chem. 1979 Feb 10;254(3):796-803
PMID: 368060
-
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):4727-30
PMID: 8197125
-
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):7424-8
PMID: 2217173
-
Isolation and characterization of Saccharomyces cerevisiae mutants resistant to Calcofluor white.
J Bacteriol. 1988 Apr;170(4):1950-4
PMID: 3280554
-
Chitin synthetase 2, a presumptive participant in septum formation in Saccharomyces cerevisiae.
J Biol Chem. 1986 Nov 15;261(32):15147-52
PMID: 2945823
-
The S. cerevisiae structural gene for chitin synthase is not required for chitin synthesis in vivo.
Cell. 1986 Jul 18;46(2):213-25
PMID: 2941152
-
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):1764-76
PMID: 1532231
-
CAL1, a gene required for activity of chitin synthase 3 in Saccharomyces cerevisiae.
J Cell Biol. 1991 Jul;114(1):101-9
PMID: 2050737
-
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):12356-60
PMID: 2061313
-
Endochitinase, a mannan-associated enzyme from Saccharomyces cerevisiae.
J Biol Chem. 1982 Feb 10;257(3):1392-7
PMID: 6799506
-
Chitinase is required for cell separation during growth of Saccharomyces cerevisiae.
J Biol Chem. 1991 Oct 15;266(29):19758-67
PMID: 1918080
-
Lariat structures are in vivo intermediates in yeast pre-mRNA splicing.
Cell. 1984 Dec;39(3 Pt 2):611-21
PMID: 6096014
-
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):14966-72
PMID: 6238967
-
Universal control mechanism regulating onset of M-phase.
Nature. 1990 Apr 5;344(6266):503-8
PMID: 2138713
-
Protein measurement with the Folin phenol reagent.
J Biol Chem. 1951 Nov;193(1):265-75
PMID: 14907713
-
Differential inhibition of chitin synthetases 1 and 2 from Saccharomyces cerevisiae by polyoxin D and nikkomycins.
Antimicrob Agents Chemother. 1991 Jan;35(1):170-3
PMID: 2014972
-
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):389-94
PMID: 6369073
-
Chemical composition of the yeast ascospore wall. The second outer layer consists of chitosan.
J Biol Chem. 1988 Aug 15;263(23):11569-74
PMID: 3042773
-
Solubilization and partial purification of yeast chitin synthetase. Confirmation of the zymogenic nature of the enzyme.
J Biol Chem. 1978 Jun 25;253(12):4419-25
PMID: 350873
-
A yeast operator overlaps an upstream activation site.
Cell. 1987 Jul 31;50(3):369-77
PMID: 3301002
-
Two chitin synthases in Saccharomyces cerevisiae.
J Biol Chem. 1987 Apr 25;262(12):5732-9
PMID: 2952643
-
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):1089-99
PMID: 1293886
-
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):645-9
PMID: 16592617
-
Hormone-induced expression of the CHS1 gene from Saccharomyces cerevisiae.
Eur J Biochem. 1989 Apr 15;181(1):243-7
PMID: 2523800
-
The use of divalent cations and pH for the determination of specific yeast chitin synthetases.
Anal Biochem. 1994 Jun;219(2):368-72
PMID: 8080094
-
Transformation of intact yeast cells treated with alkali cations.
J Bacteriol. 1983 Jan;153(1):163-8
PMID: 6336730