Abstract

We propose to investigate the role of intracellular proteinases in the metabolism and differentiation of yeast cells. Mutants defective in known proteinases or in proteolytic processes have been and will be isolated and characterized. The effects of deficiency for the proteinase or proteolytic system on the biological and biochemical characteristics of the cells will be determined. Cellular processes to be scrutinized include cell growth and division, meiosis and sporulation, and protein degradation induced by starvation, sporulation or medium change. Mutants unable to degrade unstable proteins will be characterized for the spectrum of proteins rendered stable by the mutations (nonsense fragments, thermolabile enzymes, gluconeogenic enzymes upon medium shifts, etc.) The pep4-3 mutant will be examined further to see whether larger, inactive forms (possible precursors) of vacuolar enzymes accumulate in the mutant as has been found for proteinase C (carboxypeptidase Y). Similar studies will be conducted on a mutant of a new complementation group with properties similar to pep4-3.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK018090-13
Application #
3225933
Study Section
Genetics Study Section (GEN)
Project Start
1977-08-01
Project End
1987-07-31
Budget Start
1986-08-01
Budget End
1987-07-31
Support Year
13
Fiscal Year
1986
Total Cost
Indirect Cost

  Institution

Name
Carnegie-Mellon University
Department
Type
Schools of Arts and Sciences
DUNS #
052184116
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213

  Publications

Naik, R R; Jones, E W (1998) The PBN1 gene of Saccharomyces cerevisiae: an essential gene that is required for the post-translational processing of the protease B precursor. Genetics 149:1277-92
Naik, R R; Nebes, V; Jones, E W (1997) Regulation of the proteinase B structural gene PRB1 in Saccharomyces cerevisiae. J Bacteriol 179:1469-74
Manolson, M F; Wu, B; Proteau, D et al. (1994) STV1 gene encodes functional homologue of 95-kDa yeast vacuolar H(+)-ATPase subunit Vph1p. J Biol Chem 269:14064-74
Bachhawat, A K; Manolson, M F; Murdock, D G et al. (1993) The VPH2 gene encodes a 25 kDa protein required for activity of the yeast vacuolar H(+)-ATPase. Yeast 9:175-84
Woolford, C A; Noble, J A; Garman, J D et al. (1993) Phenotypic analysis of proteinase A mutants. Implications for autoactivation and the maturation pathway of the vacuolar hydrolases of Saccharomyces cerevisiae. J Biol Chem 268:8990-8
Manolson, M F; Proteau, D; Jones, E W (1992) Evidence for a conserved 95-120 kDa subunit associated with and essential for activity of V-ATPases. J Exp Biol 172:105-12
Nebes, V L; Jones, E W (1992) N-linked glycosylation of proteinase B precursors of the yeast Saccharomyces cerevisiae is not required for proper targeting or processing of the enzyme. Yeast 8:353-9
Preston, R A; Reinagel, P S; Jones, E W (1992) Genes required for vacuolar acidity in Saccharomyces cerevisiae. Genetics 131:551-8
Manolson, M F; Proteau, D; Preston, R A et al. (1992) The VPH1 gene encodes a 95-kDa integral membrane polypeptide required for in vivo assembly and activity of the yeast vacuolar H(+)-ATPase. J Biol Chem 267:14294-303
Preston, R A; Manolson, M F; Becherer, K et al. (1991) Isolation and characterization of PEP3, a gene required for vacuolar biogenesis in Saccharomyces cerevisiae. Mol Cell Biol 11:5801-12

Showing the most recent 10 out of 17 publications