Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1982 Apr;79(7):2157–2161. doi: 10.1073/pnas.79.7.2157

Acid phosphatase polypeptides in Saccharomyces cerevisiae are encoded by a differentially regulated multigene family.

D T Rogers, J M Lemire, K A Bostian
PMCID: PMC346149  PMID: 6212932

Abstract

Two clones from a lambda phage collection containing yeast genes regulated by inorganic phosphate were shown by low-stringency hybridization to select three mRNAs that direct the in vitro synthesis of repressible acid phosphatase (EC 3.1.3.2) polypeptides p60, p58, and p56. By higher stringency hybridization one yeast fragment [8 kilobases (kb)] selects p60 mRNA and the other (5 kb) selects p56 mRNA. These EcoRI digestion fragments were subcloned in yeast transformation vectors and hybridization selection assignments were confirmed by measuring enzyme and mRNA levels in transformants. Enzyme and mRNA levels in (8-kb) high copy number transformants grown in high inorganic phosphate medium revealed a hitherto undetected acid phosphatase protein, P57, which is believed to correspond to the constitutive enzyme encoded by PHO3. The identify of the 8-kb fragment purported to contain the PHO5/PHO3 genes was confirmed by genetic mapping of an integrated copy of this fragment. The site of integration of the 5-kb fragment was demonstrated to be unlinked to the PHO5/PHO3 genes.

Full text

PDF
2157

Images in this article

Selected References

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

  1. Alwine J. C., Kemp D. J., Parker B. A., Reiser J., Renart J., Stark G. R., Wahl G. M. Detection of specific RNAs or specific fragments of DNA by fractionation in gels and transfer to diazobenzyloxymethyl paper. Methods Enzymol. 1979;68:220–242. doi: 10.1016/0076-6879(79)68017-5. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Botstein D., Falco S. C., Stewart S. E., Brennan M., Scherer S., Stinchcomb D. T., Struhl K., Davis R. W. Sterile host yeasts (SHY): a eukaryotic system of biological containment for recombinant DNA experiments. Gene. 1979 Dec;8(1):17–24. doi: 10.1016/0378-1119(79)90004-0. [DOI] [PubMed] [Google Scholar]
  4. Hereford L. M., Rosbash M. Number and distribution of polyadenylated RNA sequences in yeast. Cell. 1977 Mar;10(3):453–462. doi: 10.1016/0092-8674(77)90032-0. [DOI] [PubMed] [Google Scholar]
  5. Hinnen A., Hicks J. B., Fink G. R. Transformation of yeast. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1929–1933. doi: 10.1073/pnas.75.4.1929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hopper J. E., Bostian K. A., Rowe L. B., Tipper D. J. Translation of the L-species dsRNA genome of the killer-associated virus-like particles of Saccharomyces cerevisiae. J Biol Chem. 1977 Dec 25;252(24):9010–9017. [PubMed] [Google Scholar]
  7. Kaneko Y., Toh-e A., Oshima Y. Identification of the genetic locus for the structural gene and a new regulatory gene for the synthesis of repressible alkaline phosphatase in Saccharomyces cerevisiae. Mol Cell Biol. 1982 Feb;2(2):127–137. doi: 10.1128/mcb.2.2.127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Klar A. J., Halvorson H. O. Effect of GAL4 gene dosage on the level of galactose catabolic enzymes in Saccharomyces cerevisiae. J Bacteriol. 1976 Jan;125(1):379–381. doi: 10.1128/jb.125.1.379-381.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kramer R. A., Andersen N. Isolation of yeast genes with mRNA levels controlled by phosphate concentration. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6541–6545. doi: 10.1073/pnas.77.11.6541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Maniatis T., Jeffrey A., Kleid D. G. Nucleotide sequence of the rightward operator of phage lambda. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1184–1188. doi: 10.1073/pnas.72.3.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Mildner P., Barbarić S., Golubić Z., Ries B. Purification of protoplast-secreted acid phosphatase from baker's yeast. Effect on adenosine triphosphatase activity. Biochim Biophys Acta. 1976 Mar 11;429(1):274–282. doi: 10.1016/0005-2744(76)90050-4. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Schurr A., Yagil E. Regulation and characterization of acid and alkaline phosphatase in yeast. J Gen Microbiol. 1971 Mar;65(3):291–303. doi: 10.1099/00221287-65-3-291. [DOI] [PubMed] [Google Scholar]
  15. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. 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]
  18. 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]
  19. Toh-e A., Kakimoto S. Genes coding for the structure of the acid phosphatases in Saccharomyces cerevisiae. Mol Gen Genet. 1975 Dec 30;143(1):65–70. doi: 10.1007/BF00269421. [DOI] [PubMed] [Google Scholar]
  20. Toh-e A., Kakimoto S., Oshima Y. Two new genes controlling the constitutive acid phosphatase synthesis in Saccharomyces cerevisiae. Mol Gen Genet. 1975 Nov 3;141(1):81–83. doi: 10.1007/BF00332380. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Van Rijn H. J., Boer P., Steyn-Parvé E. P. Biosynthesis of acid phosphatase of baker's yeast. Factors influencing its production by protoplasts and characterization of the secreted enzyme. Biochim Biophys Acta. 1972 May 12;268(2):431–441. doi: 10.1016/0005-2744(72)90339-7. [DOI] [PubMed] [Google Scholar]
  23. Woolford J. L., Jr, Rosbash M. The use of R-looping for structural gene identification and mRNA purification. Nucleic Acids Res. 1979 Jun 11;6(7):2483–2497. doi: 10.1093/nar/6.7.2483. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES