Abstract

The goal is to elucidate the structures functional roles and physiological regulation of the expression of multiple forms of type II cAMP-dependent protein kinases (PKII). We have shown that the basis for PKII diversity is the tissue-specific expression of several distinct cAMP-binding or regulatory (RII) subunits. Cloned cDNA sequences corresponding to mRNA for a major cerebral cortex RII isoform (RII-B) will be isolated from a Lambda gt11 library. Th RII-B cDNA probe will be used to obtain cDNA clones for RII subunit isoforms produced in non-neural bovine tissues (RII-H) and murine erythroleukemic cells (RII-52 and RII-54) from Lamba gt11 or pU C18 cDNA libraries. The number and sizes of mRNAs coding for RII subunits in adult tissues will be determined. Northern gels and dot-blot analyses will be used to quantify changes in RII mRNA content during the induction of RII-52 by cAMP treatment and erythroid differentiation. The possibility that regulation of RII-52 biosynthesis is exerted at the levels of transcription, processing and/or mRNA stabilization will be examined. The degree to which the RII isoforms are homologous and divergent will be investigated by RNAse mapping and primer extension analysis, and subsequently, direct sequencing using the Sanger chain termination procedure. cDNA probes will be used to isolate cloned genomic sequences for the RII isoforms. THe organization of RII genes will be established by restriction and S1 mapping and sequencing 1 kb upstream from the 5' end of the structural gene to begin to assess the DNA sequences involved in the differential tissue-specific, developmental and hormone stimulated expression of RII isoforms. Finally, P75, a calmodulin binding protein that complexes RII-B, will be purified to homogeneity and characterized by hydrodynamic methods. Binding and interactions among P75, calmodulin and RII-B will be quantitatively analyzed and effects of phospho/dephospho-P75 on the functional properties of PKII-B will be determined.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM022792-12
Application #
3271335
Study Section
Physiological Chemistry Study Section (PC)
Project Start
1976-03-01
Project End
1991-03-30
Budget Start
1989-07-01
Budget End
1990-06-30
Support Year
12
Fiscal Year
1989
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

Han, Ping; Sonati, Pushpalatha; Rubin, Charles et al. (2006) PDE7A1, a cAMP-specific phosphodiesterase, inhibits cAMP-dependent protein kinase by a direct interaction with C. J Biol Chem 281:15050-7
Zhang, L; Wu, S L; Rubin, C S (2001) Structural properties and mechanisms that govern association of C kinase adapter 1 with protein kinase C3 and the cell periphery. J Biol Chem 276:10476-84
Zhang, L; Wu, S L; Rubin, C S (2001) A novel adapter protein employs a phosphotyrosine binding domain and exceptionally basic N-terminal domains to capture and localize an atypical protein kinase C: characterization of Caenorhabditis elegans C kinase adapter 1, a protein that avidly binds pr J Biol Chem 276:10463-75
Rossi, E A; Li, Z; Feng, H et al. (1999) Characterization of the targeting, binding, and phosphorylation site domains of an A kinase anchor protein and a myristoylated alanine-rich C kinase substrate-like analog that are encoded by a single gene. J Biol Chem 274:27201-10
Davare, M A; Dong, F; Rubin, C S et al. (1999) The A-kinase anchor protein MAP2B and cAMP-dependent protein kinase are associated with class C L-type calcium channels in neurons. J Biol Chem 274:30280-7
Feliciello, A; Cardone, L; Garbi, C et al. (1999) Yotiao protein, a ligand for the NMDA receptor, binds and targets cAMP-dependent protein kinase II(1). FEBS Lett 464:174-8
Li, Z; Rossi, E A; Hoheisel, J D et al. (1999) Generation of a novel A kinase anchor protein and a myristoylated alanine-rich C kinase substrate-like analog from a single gene. J Biol Chem 274:27191-200
Angelo, R; Rubin, C S (1998) Molecular characterization of an anchor protein (AKAPCE) that binds the RI subunit (RCE) of type I protein kinase A from Caenorhabditis elegans. J Biol Chem 273:14633-43
Feliciello, A; Rubin, C S; Avvedimento, E V et al. (1998) Expression of a kinase anchor protein 121 is regulated by hormones in thyroid and testicular germ cells. J Biol Chem 273:23361-6
Dong, F; Feldmesser, M; Casadevall, A et al. (1998) Molecular characterization of a cDNA that encodes six isoforms of a novel murine A kinase anchor protein. J Biol Chem 273:6533-41

Showing the most recent 10 out of 36 publications