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.
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