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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
. 1983 Jan;80(2):510–514. doi: 10.1073/pnas.80.2.510

Mutations in PEP4 locus of Saccharomyces cerevisiae block final step in maturation of two vacuolar hydrolases.

G S Zubenko, F J Park, E W Jones
PMCID: PMC393408  PMID: 6340101

Abstract

The biosynthesis of carboxypeptidase Y (EC 3.4.16.1) and proteinase A (EC 3.4.23.6) in yeast cells involves a series of posttranslational events, the last of which is dependent upon a function supplied by the PEP4 gene. Because pep4 mutations result in a 90-95% reduction in the levels of activity of at least three additional vacuolar hydrolases, it is likely that these, and perhaps all, yeast vacuolar hydrolases are synthesized as inactive precursors, which mature by a common mechanism that depends upon a function supplied by the PEP4 gene. The pep4 mutation shows an apparent gene dosage effect on levels of activity of proteinases A and B but not on the level of activity of carboxypeptidase Y. This effect appears to come about because the maturation machinery is capable of discriminating among these hydrolase precursors.

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Selected References

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  1. Czichi U., Lennarz W. J. Localization of the enzyme system for glycosylation of proteins via the lipid-linked pathway in rough endoplasmic reticulum. J Biol Chem. 1977 Nov 25;252(22):7901–7904. [PubMed] [Google Scholar]
  2. Esmon B., Novick P., Schekman R. Compartmentalized assembly of oligosaccharides on exported glycoproteins in yeast. Cell. 1981 Aug;25(2):451–460. doi: 10.1016/0092-8674(81)90063-5. [DOI] [PubMed] [Google Scholar]
  3. Hasilik A., Neufeld E. F. Biosynthesis of lysosomal enzymes in fibroblasts. Phosphorylation of mannose residues. J Biol Chem. 1980 May 25;255(10):4946–4950. [PubMed] [Google Scholar]
  4. Hasilik A., Tanner W. Biosynthesis of carboxypeptidase Y in yeast. Evidence for a precursor form of the glycoprotein. Biochem Biophys Res Commun. 1976 Oct 18;72(4):1430–1436. doi: 10.1016/s0006-291x(76)80173-8. [DOI] [PubMed] [Google Scholar]
  5. Hasilik A., Tanner W. Biosynthesis of the vacuolar yeast glycoprotein carboxypeptidase Y. Conversion of precursor into the enzyme. Eur J Biochem. 1978 Apr 17;85(2):599–608. doi: 10.1111/j.1432-1033.1978.tb12275.x. [DOI] [PubMed] [Google Scholar]
  6. Hasilik A., Tanner W. Carbohydrate moiety of carboxypeptidase Y and perturbation of its biosynthesis. Eur J Biochem. 1978 Nov 15;91(2):567–575. doi: 10.1111/j.1432-1033.1978.tb12710.x. [DOI] [PubMed] [Google Scholar]
  7. Hasilik A., Tanner W. Inhibition of the apparent rate of synthesis on the vacuolar glycoprotein carboxypeptidase Y and its protein antigen by turicamycin in Saccharomyces cerevisiae. Antimicrob Agents Chemother. 1976 Sep;10(3):402–410. doi: 10.1128/aac.10.3.402. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hayashi R., Aibara S., Hata T. A unique carboxypeptidase activity of yeast proteinase C. Biochim Biophys Acta. 1970 Aug 15;212(2):359–361. doi: 10.1016/0005-2744(70)90218-4. [DOI] [PubMed] [Google Scholar]
  9. Hayashi R., Moore S., Stein W. H. Carboxypeptidase from yeast. Large scale preparation and the application to COOH-terminal analysis of peptides and proteins. J Biol Chem. 1973 Apr 10;248(7):2296–2302. [PubMed] [Google Scholar]
  10. Hemmings B. A., Zubenko G. S., Hasilik A., Jones E. W. Mutant defective in processing of an enzyme located in the lysosome-like vacuole of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1981 Jan;78(1):435–439. doi: 10.1073/pnas.78.1.435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hinze H., Betz H., Saheki T., Holzer H. Formation of a complex between yeast proteinases A and B. Hoppe Seylers Z Physiol Chem. 1975 Aug;356(8):1259–1264. doi: 10.1515/bchm2.1975.356.2.1259. [DOI] [PubMed] [Google Scholar]
  12. Jones E. W. Proteinase mutants of Saccharomyces cerevisiae. Genetics. 1977 Jan;85(1):23–33. doi: 10.1093/genetics/85.1.23. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jones E. W., Zubenko G. S., Parker R. R. PEP4 gene function is required for expression of several vacuolar hydrolases in Saccharomyces cerevisiae. Genetics. 1982 Dec;102(4):665–677. doi: 10.1093/genetics/102.4.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Kiely M. L., McKnight G. S., Schimke R. T. Studies on the attachment of carbohydrate to ovalbumin nascent chains in hen oviduct. J Biol Chem. 1976 Sep 25;251(18):5490–5495. [PubMed] [Google Scholar]
  16. Kuhn R. W., Walsh K. A., Neurath H. Isolation and partial characterization of an acid carboxypeptidase from yeast. Biochemistry. 1974 Sep 10;13(19):3871–3877. doi: 10.1021/bi00716a008. [DOI] [PubMed] [Google Scholar]
  17. Kuo S. C., Lampen J. O. Tunicamycin--an inhibitor of yeast glycoprotein synthesis. Biochem Biophys Res Commun. 1974 May 7;58(1):287–295. doi: 10.1016/0006-291x(74)90925-5. [DOI] [PubMed] [Google Scholar]
  18. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  19. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  20. Lehle L., Bauer F., Tanner W. The formation of glycosidic bonds in yeast glycoproteins. Intracellular localisation of the reactions. Arch Microbiol. 1977 Jul 26;114(1):77–81. doi: 10.1007/BF00429634. [DOI] [PubMed] [Google Scholar]
  21. Lehle L. Biosynthesis of the core region of yeast mannoproteins. Formation of a glucosylated dolichol-bound oligosaccharide precursor, its transfer to protein and subsequent modification. Eur J Biochem. 1980 Aug;109(2):589–601. doi: 10.1111/j.1432-1033.1980.tb04832.x. [DOI] [PubMed] [Google Scholar]
  22. Mechler B., Müller M., Müller H., Meussdoerffer F., Wolf D. H. In vivo biosynthesis of the vacuolar proteinases A and B in the yeast Saccharomyces cerevisiae. J Biol Chem. 1982 Oct 10;257(19):11203–11206. [PubMed] [Google Scholar]
  23. Novick P., Ferro S., Schekman R. Order of events in the yeast secretory pathway. Cell. 1981 Aug;25(2):461–469. doi: 10.1016/0092-8674(81)90064-7. [DOI] [PubMed] [Google Scholar]
  24. Roberts J. W., Roberts C. W. Proteolytic cleavage of bacteriophage lambda repressor in induction. Proc Natl Acad Sci U S A. 1975 Jan;72(1):147–151. doi: 10.1073/pnas.72.1.147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Schachter H., Jabbal I., Hudgin R. L., Pinteric L., McGuire E. J., Roseman S. Intracellular localization of liver sugar nucleotide glycoprotein glycosyltransferases in a Golgi-rich fraction. J Biol Chem. 1970 Mar 10;245(5):1090–1100. [PubMed] [Google Scholar]
  26. Stevens T., Esmon B., Schekman R. Early stages in the yeast secretory pathway are required for transport of carboxypeptidase Y to the vacuole. Cell. 1982 Sep;30(2):439–448. doi: 10.1016/0092-8674(82)90241-0. [DOI] [PubMed] [Google Scholar]
  27. Tarentino A. L., Maley F. Purification and properties of an endo-beta-N-acetylglucosaminidase from Streptomyces griseus. J Biol Chem. 1974 Feb 10;249(3):811–817. [PubMed] [Google Scholar]
  28. Tarentino A. L., Plummer T. H., Jr, Maley F. The release of intact oligosaccharides from specific glycoproteins by endo-beta-N-acetylglucosaminidase H. J Biol Chem. 1974 Feb 10;249(3):818–824. [PubMed] [Google Scholar]
  29. Trimble R. B., Maley F. The use of endo-beta-N-acetylglucosaminidase H in characterizing the structure and function of glycoproteins. Biochem Biophys Res Commun. 1977 Oct 10;78(3):935–944. doi: 10.1016/0006-291x(77)90512-5. [DOI] [PubMed] [Google Scholar]
  30. Whur P., Herscovics A., Leblond C. P. Radioautographic visualization of the incorporation of galactose-3H and mannose-3H by rat thyroids in vitro in relation to the stages of thyroglobulin synthesis. J Cell Biol. 1969 Nov;43(2):289–311. doi: 10.1083/jcb.43.2.289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wolf D. H., Weiser U. Studies on a carboxypeptidase Y mutant of yeast and evidence for a second carboxypeptidase Activity. Eur J Biochem. 1977 Mar 1;73(2):553–556. doi: 10.1111/j.1432-1033.1977.tb11350.x. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Zubenko G. S., Park F. J., Jones E. W. Genetic properties of mutations at the PEP4 locus in Saccharomyces cerevisiae. Genetics. 1982 Dec;102(4):679–690. doi: 10.1093/genetics/102.4.679. [DOI] [PMC free article] [PubMed] [Google Scholar]

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