Abstract

The research will continue the characterization of the physical structure and regulation of several genes whose expression is coordinately regulated by cAMP during Dictyostelium development. The first of these genes is the cAMP phosphodiesterase, which we have cloned. This gene will be further characterized and its regulation studied. The second gene is an inhibitor of the cAMP phosphodiesterase and is repressed by cAMP. The inhibitor glycoprotein has been purified and the amino terminal sequence has been determined. The gene coding for this glycoprotein will be cloned by the same techniques used for the cAMP phosphodiesterase gene. A third protein, the cAMP protein kinase will be studied in collaboration with another laboratory and compared to the first two genes. The cAMP dependent protein kinase regulatory and catalytic subunit genes will be recovered from a cDNA library prepared with cAMP induced mRNA and introduced into the lambda vector gt11. Several mutants which lack cAMP phosphodiesterase will be analysed to determine if they are structural or regulatory defects. The role of cAMP dependent protein kinase in the regulation of the other genes will be analysed. The cAMP phosphodiesterase gene will be manipulated in a way which allows its expression and use to degrade cAMP in Dictyostelium and other cells. This should provide a new tool for the study of cAMP metabolism.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM033136-04
Application #
3282463
Study Section
Genetics Study Section (GEN)
Project Start
1983-07-01
Project End
1991-02-28
Budget Start
1986-03-01
Budget End
1987-02-28
Support Year
4
Fiscal Year
1986
Total Cost
Indirect Cost

  Institution

Name
Columbia University (N.Y.)
Department
Type
Schools of Medicine
DUNS #
064931884
City
New York
State
NY
Country
United States
Zip Code
10027

  Publications

Tekinay, Turgay; Wu, Mary Y; Otto, Grant P et al. (2006) Function of the Dictyostelium discoideum Atg1 kinase during autophagy and development. Eukaryot Cell 5:1797-806
Otto, Grant P; Wu, Mary Y; Clarke, Margaret et al. (2004) Macroautophagy is dispensable for intracellular replication of Legionella pneumophila in Dictyostelium discoideum. Mol Microbiol 51:63-72
Otto, Grant P; Wu, Mary Y; Kazgan, Nevzat et al. (2004) Dictyostelium macroautophagy mutants vary in the severity of their developmental defects. J Biol Chem 279:15621-9
Weening, Karin E; Wijk, Irene Verkerke-Van; Thompson, Christopher R et al. (2003) Contrasting activities of the aggregative and late PDSA promoters in Dictyostelium development. Dev Biol 255:373-82
Otto, Grant P; Wu, Mary Y; Kazgan, Nevzat et al. (2003) Macroautophagy is required for multicellular development of the social amoeba Dictyostelium discoideum. J Biol Chem 278:17636-45
Pukatzki, Stefan; Kessin, Richard H; Mekalanos, John J (2002) The human pathogen Pseudomonas aeruginosa utilizes conserved virulence pathways to infect the social amoeba Dictyostelium discoideum. Proc Natl Acad Sci U S A 99:3159-64
Ennis, H L; Dao, D N; Pukatzki, S U et al. (2000) Dictyostelium amoebae lacking an F-box protein form spores rather than stalk in chimeras with wild type. Proc Natl Acad Sci U S A 97:3292-7
Dao, D N; Kessin, R H; Ennis, H L (2000) Developmental cheating and the evolutionary biology of Dictyostelium and Myxococcus. Microbiology 146 ( Pt 7):1505-12
Pukatzki, S; Ennis, H L; Kessin, R H (2000) A genetic interaction between a ubiquitin-like protein and ubiquitin-mediated proteolysis in Dictyostelium discoideum(1). Biochim Biophys Acta 1499:154-163
Pukatzki, S; Tordilla, N; Franke, J et al. (1998) A novel component involved in ubiquitination is required for development of Dictyostelium discoideum. J Biol Chem 273:24131-8

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