Abstract
The synthesis of repressible acid phosphatase in Saccharomyces cerevisiae was examined under conditions of blocked derepression as described by Toh-e et al. (Mol. Gen. Genet. 162:139-149, 1978). Based on a genetic and biochemical analysis of the phenomenon these authors proposed a new regulatory model for acid phosphatase expression involving a simultaneous interaction of regulatory factors in the control of structural gene transcription. We demonstrate here that under growth conditions that fail to produce acid phosphatase the enzyme is readily inactivated. Furthermore, we demonstrate under these conditions the production of acid phosphatase mRNA which is active both in vitro and in vivo in the synthesis of enzyme. This eliminates any step prior to translation of acid phosphatase polypeptide as an explanation for the phenomenon. We interpret our results for the block in appearance of acid phosphatase as a result of both deaccelerated growth and cellular biosynthesis during derepression, accompanied by an enhanced instability of the enzyme.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Boer P., Steyn-Parvé E. P. Isolation and purification of an acid phosphatase from baker's yeast (Saccharomyces cerevisiae). Biochim Biophys Acta. 1966 Nov 15;128(2):400–402. doi: 10.1016/0926-6593(66)90189-5. [DOI] [PubMed] [Google Scholar]
- Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
- Bostian K. A., Hopper J. E., Rogers D. T., Tipper D. J. Translational analysis of the killer-associated virus-like particle dsRNA genome of S. cerevisiae: M dsRNA encodes toxin. Cell. 1980 Feb;19(2):403–414. doi: 10.1016/0092-8674(80)90514-0. [DOI] [PubMed] [Google Scholar]
- Bostian K. A., Lee R. C., Halvorson H. O. Preparative fractionation of nucleic acids by agarose gel electrophoresis. Anal Biochem. 1979 May;95(1):174–182. doi: 10.1016/0003-2697(79)90201-x. [DOI] [PubMed] [Google Scholar]
- Bostian K. A., Lemire J. M., Cannon L. E., Halvorson H. O. In vitro synthesis of repressible yeast acid phosphatase: identification of multiple mRNAs and products. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4504–4508. doi: 10.1073/pnas.77.8.4504. [DOI] [PMC free article] [PubMed] [Google Scholar]
- HEREDIA C. F., YEN F., SOLS A. Role and formation of the acid phosphatase in yeast. Biochem Biophys Res Commun. 1963 Jan 18;10:14–18. doi: 10.1016/0006-291x(63)90259-6. [DOI] [PubMed] [Google Scholar]
- Hopper J. E., Broach J. R., Rowe L. B. Regulation of the galactose pathway in Saccharomyces cerevisiae: induction of uridyl transferase mRNA and dependency on GAL4 gene function. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2878–2882. doi: 10.1073/pnas.75.6.2878. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Matsumoto K., Toh-e A., Oshima Y. Genetic control of galactokinase synthesis in Saccharomyces cerevisiae: evidence for constitutive expression of the positive regulatory gene gal4. J Bacteriol. 1978 May;134(2):446–457. doi: 10.1128/jb.134.2.446-457.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Perlman D., Hopper J. E. Constitutive synthesis of the GAL4 protein, a galactose pathway regulator in Saccharomyces cerevisiae. Cell. 1979 Jan;16(1):89–95. doi: 10.1016/0092-8674(79)90190-9. [DOI] [PubMed] [Google Scholar]
- Rothman J. E., Katz F. N., Lodish H. F. Glycosylation of a membrane protein is restricted to the growing polypeptide chain but is not necessary for insertion as a transmembrane protein. Cell. 1978 Dec;15(4):1447–1454. doi: 10.1016/0092-8674(78)90068-5. [DOI] [PubMed] [Google Scholar]
- SCHMIDT G., BARTSCH G., LAUMONT M. C., HERMAN T., LISS M. Acid phosphatase of bakers' yeast: an enzyme of the external cell surface. Biochemistry. 1963 Jan-Feb;2:126–131. doi: 10.1021/bi00901a022. [DOI] [PubMed] [Google Scholar]
- Studier F. W. Analysis of bacteriophage T7 early RNAs and proteins on slab gels. J Mol Biol. 1973 Sep 15;79(2):237–248. doi: 10.1016/0022-2836(73)90003-x. [DOI] [PubMed] [Google Scholar]
- To-E A., Ueda Y., Kakimoto S. I., Oshima Y. Isolation and characterization of acid phosphatase mutants in Saccharomyces cerevisiae. J Bacteriol. 1973 Feb;113(2):727–738. doi: 10.1128/jb.113.2.727-738.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Toh-E A., Oshima Y. Characterization of a dominant, constitutive mutation, PHOO, for the repressible acid phosphatase synthesis in Saccharomyces cerevisiae. J Bacteriol. 1974 Nov;120(2):608–617. doi: 10.1128/jb.120.2.608-617.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Toh-e A., Inouye S., Oshima Y. Structure and function of the PHO82-pho4 locus controlling the synthesis of repressible acid phosphatase of Saccharomyces cerevisiae. J Bacteriol. 1981 Jan;145(1):221–232. doi: 10.1128/jb.145.1.221-232.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Toh-e A., Kobayashi S., Oshima Y. Disturbance of the machinery for the gene expression by acidic pH in the repressible acid phosphatase system of Saccharomyces cerevisiae. Mol Gen Genet. 1978 Jun 14;162(2):139–149. doi: 10.1007/BF00267870. [DOI] [PubMed] [Google Scholar]
- Ueda Y., To-E A., Oshima Y. Isolation and characterization of recessive, constitutive mutations for repressible acid phosphatase synthesis in Saccharomyces cerevisiae. J Bacteriol. 1975 Jun;122(3):911–922. doi: 10.1128/jb.122.3.911-922.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]