The glycolytic pathway is quantitatively the chief metabolic route of most cells, normal and diseased. Although differing in details between higher and lower cells, it is the same pathway and the same biochemistry in microbes and presents, in them as in higher cells, avariety of matters still being clarified. These include questions of isoenzyme function, allosteric controls, glucose uptake, the possibility of an enzyme complex, and the issue of how flux is determined in vivo, as well as description of its genetics. Our work, with Saccharomyces cerevisiae and Escherichia coli, touches on most of these points. The current focus is on the hexokinases and their roles in glucose uptake (in yeast), the fructose-6- P/fructose-1,6-P2 interconversion (in both organisms), and the aldolases (in E. coli), and several projects, including the latter, concern phosphorylation of enzymes and its mutational alteration. We also propose work on the putative glycolytic complex, and some new lines relating to the description of metabolism as a whole, which derive from having a variety of strains with high level or altered enzymes. These include experiments on rate limitation in the pathway and on transit time.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM021098-15
Application #
3270242
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1977-08-01
Project End
1993-07-31
Budget Start
1988-08-01
Budget End
1989-07-31
Support Year
15
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Harvard University
Department
Type
Schools of Medicine
DUNS #
082359691
City
Boston
State
MA
Country
United States
Zip Code
02115
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Uemura, H; Jigami, Y (1992) GCR3 encodes an acidic protein that is required for expression of glycolytic genes in Saccharomyces cerevisiae. J Bacteriol 174:5526-32
Paravicini, G; Kretschmer, M (1992) The yeast FBP26 gene codes for a fructose-2,6-bisphosphatase. Biochemistry 31:7126-33
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Kretschmer, M; Fraenkel, D G (1991) Yeast 6-phosphofructo-2-kinase: sequence and mutant. Biochemistry 30:10663-72
Vojtek, A B; Fraenkel, D G (1990) Phosphorylation of yeast hexokinases. Eur J Biochem 190:371-5
Uemura, H; Fraenkel, D G (1990) gcr2, a new mutation affecting glycolytic gene expression in Saccharomyces cerevisiae. Mol Cell Biol 10:6389-96
Babul, J; Fraenkel, D G (1988) Phosphate modification of fructose-1,6-bisphosphate aldolase in Escherichia coli. Biochem Biophys Res Commun 151:1033-8
Sedivy, J M; Babul, J; Fraenkel, D G (1986) AMP-insensitive fructose bisphosphatase in Escherichia coli and its consequences. Proc Natl Acad Sci U S A 83:1656-9
Baker, H V (1986) Glycolytic gene expression in Saccharomyces cerevisiae: nucleotide sequence of GCR1, null mutants, and evidence for expression. Mol Cell Biol 6:3774-84

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