Phosphatidylinositol metabolism is important to the growth of eukaryotic organisms. The agonist-stimulated hydrolysis of membrane polyphosphoinositides in animals result in the stimulation of various cellular responses. The enzyme that catalyzes the initial step in the synthesis of the polyphosphoinositides is phosphatidylinositol kinase. This application will focus on the regulation of phosphatidylinositol kinase in the simple eukaryote, Saccharomyces cerevisiae. Phosphatidylinositol kinase will be purified to homogeneity and will be characterized with respect to enzymological, chemical, and physical properties. The purified enzyme will be reconstituted into well defined unilamellar phospholipid vesicles to study the regulation of activity. Regulation will be studied by altering the composition and environment of the vesicles and by possible phosphorylation and dephosphorylation of the enzyme. Antibodies specific for phosphatidylinositol kinase will be prepared and used as a probe to study the regulation of phosphatidylinositol kinase formation in cells grown under various conditions. The proposed studies should provide insight into the regulation of polyphosphoinositide synthesis. The results found with S. cerevisiae should be relevant to higher eukaryotic organisms.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM035655-03
Application #
3288633
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1986-07-01
Project End
1989-06-30
Budget Start
1988-07-01
Budget End
1989-06-30
Support Year
3
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Rutgers University
Department
Type
Earth Sciences/Resources
DUNS #
038633251
City
New Brunswick
State
NJ
Country
United States
Zip Code
08901
Carman, G M; Zeimetz, G M (1996) Regulation of phospholipid biosynthesis in the yeast Saccharomyces cerevisiae. J Biol Chem 271:13293-6
Carman, G M; Deems, R A; Dennis, E A (1995) Lipid signaling enzymes and surface dilution kinetics. J Biol Chem 270:18711-4
Nickels Jr, J T; Buxeda, R J; Carman, G M (1994) Regulation of phosphatidylinositol 4-kinase from the yeast Saccharomyces cerevisiae by CDP-diacylglycerol. J Biol Chem 269:11018-24
Nickels Jr, J T; Carman, G M (1993) Photoaffinity labeling of the 45-kDa and 55-kDa forms of phosphatidylinositol 4-kinase from the yeast Saccharomyces cerevisiae. J Biol Chem 268:24083-8
Buxeda, R J; Nickels Jr, J T; Carman, G M (1993) Regulation of the 45- and 55-kDa forms of phosphatidylinositol 4-kinase from the yeast Saccharomyces cerevisiae by nucleotides. J Biol Chem 268:6248-55
Carman, G M; Belunis, C J; Nickels Jr, J T (1992) Phosphatidylinositol 4-kinase from yeast. Methods Enzymol 209:183-9
Nickels Jr, J T; Buxeda, R J; Carman, G M (1992) Purification, characterization, and kinetic analysis of a 55-kDa form of phosphatidylinositol 4-kinase from Saccharomyces cerevisiae. J Biol Chem 267:16297-304
Buxeda, R J; Nickels Jr, J T; Belunis, C J et al. (1991) Phosphatidylinositol 4-kinase from Saccharomyces cerevisiae. Kinetic analysis using Triton X-100/phosphatidylinositol-mixed micelles. J Biol Chem 266:13859-65
Carman, G M; Henry, S A (1989) Phospholipid biosynthesis in yeast. Annu Rev Biochem 58:635-69
Belunis, C J; Bae-Lee, M; Kelley, M J et al. (1988) Purification and characterization of phosphatidylinositol kinase from Saccharomyces cerevisiae. J Biol Chem 263:18897-903

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