Abstract

The proposed studies are designed to investigate the molecular mechanism(s) controlling the expression of the structural genes involved in maltose (and starch) metabolism in yeast. We are particularly interested in characterizing the cis and trans regulatory components of the MAL locus involved in induction by maltose and carbon catabolite repression by glucose. Effort is directed at the structure and function of the maltase and maltose permease promoter elements and of the trans-acting regulatory protein and its site(s) of action. 1. Complete the sequencing of the MAL6 locus, characterize the MALR repeated sequence (MALRR) and sequence the MAL1 intergenic (IG) region. In these experiments, the sequencing of the entire MAL6 locus and of a repeated sequence (MAL6RR) homologous to MAL6R will be completed. Preliminary studies on the properties of MAL6RR will be initiated. 2. Additional studies on the promoter elements in the MAL6 IG region. The UASM sequence will be studied in greater detail with respect to its boundaries, whether its orientation and distance from the TATA sequence are important and for its possible role in regulating the divergently transcribed permease gene. The role of the T-rich sequence in glucose repression will be studied further and examined to determine whether specific proteins bind to it. 3. Studies on the MAL6R encoded regulatory protein. The regulatory protein will be expressed in E. coli and synthesized in vitro in order to make antibodies to it. The regulatory protein and some of its mutationally altered derivatives will be studied for interaction with the maltase promoter region. 4. Search for maltase constitutive and glucose repression insensitive mutants. Constitutive mutants will be generated by mutagenizing the regulatory gene and the IG region. They will be checked for their repression by glucose. 5. Duplication of the MAL1 locus. Diploids will be constructed which will allow one to analyze random spores for duplications of the MAL1 locus. 6. Studies on yeast glucoamylase. Sequencing of the STA2 gene will be completed, the STA10 gene cloned, the STA genes mapped and the regulation of STA genes by MATa/MATalpha investigated.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM028572-09
Application #
3275830
Study Section
Molecular Biology Study Section (MBY)
Project Start
1981-03-01
Project End
1992-11-30
Budget Start
1989-12-01
Budget End
1990-11-30
Support Year
9
Fiscal Year
1990
Total Cost
Indirect Cost

  Institution

Name
Albert Einstein College of Medicine
Department
Type
Schools of Medicine
DUNS #
009095365
City
Bronx
State
NY
Country
United States
Zip Code
10461

  Publications

Lambrechts, M G; Pretorius, I S; Marmur, J et al. (1995) The S1, S2 and SGA1 ancestral genes for the STA glucoamylase genes all map to chromosome IX in Saccharomyces cerevisiae. Yeast 11:783-7
Yao, B; Sollitti, P; Zhang, X et al. (1994) Shared control of maltose induction and catabolite repression of the MAL structural genes in Saccharomyces. Mol Gen Genet 243:622-30
Sollitti, P; Merkler, D J; Estupinan, B et al. (1993) Yeast AMP deaminase. Catalytic activity in Schizosaccharomyces pombe and chromosomal location in Saccharomyces cerevisiae. J Biol Chem 268:4549-55
Yao, B; Marmur, J; Sollitti, P (1993) Construction of glucose-repressible yeast expression vectors. Gene 137:223-6
Lambrechts, M G; Pretorius, I S; Sollitti, P et al. (1991) Primary structure and regulation of a glucoamylase-encoding gene (STA2) in Saccharomyces diastaticus. Gene 100:95-103
Yao, B; Sollitti, P; Marmur, J (1989) Primary structure of the maltose-permease-encoding gene of Saccharomyces carlsbergensis. Gene 79:189-97
Vanoni, M; Sollitti, P; Goldenthal, M et al. (1989) Structure and regulation of the multigene family controlling maltose fermentation in budding yeast. Prog Nucleic Acid Res Mol Biol 37:281-322
Chow, T H; Sollitti, P; Marmur, J (1989) Structure of the multigene family of MAL loci in Saccharomyces. Mol Gen Genet 217:60-9
Sollitti, P; Marmur, J (1988) Primary structure of the regulatory gene from the MAL6 locus of Saccharomyces carlsbergensis. Mol Gen Genet 213:56-62
Pretorius, I S; Laing, E; Pretorius, G H et al. (1988) Expression of a Bacillus alpha-amylase gene in yeast. Curr Genet 14:1-8

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