The programming of cell function at the gene level during ovarian development and differentiation requires control mechanisms in which cyclic AMP and cyclic AMP-dependent protein kinase subunits are believed to be actively involved. The function of cellular regulatory proteins may be altered or modulated through the action of the catalytic subunit of cyclic AMP-dependent protein kinase which is under control by gonfadotropic hormones (FSH/LH) mediated through the action of cyclic AMP. The proposed research is designed to elucidate the molecular actions of FSH on the cyclic AMP-dependent protein kinase system in rat ovarian granulosa cells. Specifically, we aim to study the effects of nuclear cyclic AMP-dependent protein kinase on the selective phosphorylation of nuclear nonhistone proteins, histone H1 and the regulatory subunit RII. Furthermore, we aim to study the rate of biosynthesis/degradation of ovarian cyclic AMP-dependent protein kinase subunits; levels of cytoplasmic and nuclear subunits RI, RII and C. In addition, the association of RI, RII, C, histone H1 with transcriptionally active regions of ovarian chromatin will be studied by DNAase I treatment of chromatin. A new aspect of this study deals with the identification of the regulatory RII isoform which exhibits topoisomerase activity. The data will contribute toward elucidation and understanding of the factors involved in the cyclic AMP and cyclic AMP-dependent protein kinase-mediated control of ovarian differentiation and function.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD012046-09
Application #
3311768
Study Section
Biochemical Endocrinology Study Section (BCE)
Project Start
1979-03-01
Project End
1991-11-30
Budget Start
1987-12-01
Budget End
1988-11-30
Support Year
9
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Northwestern University at Chicago
Department
Type
School of Medicine & Dentistry
DUNS #
005436803
City
Chicago
State
IL
Country
United States
Zip Code
60611
Kwast-Welfeld, J; Soong, C J; Short, M L et al. (1989) Identification of rat ovarian nuclear factors that interact with the cAMP-inducible lactate dehydrogenase A subunit promoter. J Biol Chem 264:6941-7
Hand, A R; Jungmann, R A (1989) Localization of cellular regulatory proteins using postembedding immunogold labeling. Am J Anat 185:183-96
Kwast-Welfeld, J; Jungmann, R A (1988) Hormonal regulation of nuclear cyclic AMP-dependent protein kinase subunit levels in rat ovaries. J Biol Chem 263:14343-50
Jungmann, R A; Kuettel, M R; Squinto, S P et al. (1988) Using immunocolloidal gold electron microscopy to investigate cAMP-dependent protein kinase cellular compartmentalization. Methods Enzymol 159:225-35
Kuettel, M R; Squinto, S P; Kwast-Welfeld, J et al. (1985) Localization of nuclear subunits of cyclic AMP-dependent protein kinase by the immunocolloidal gold method. J Cell Biol 101:965-75