The accumulation of karyophilic proteins, molecules which are highly concentrated in the nucleus, appears to involve nuclear binding and, in some instances, facilitated transport across the envelope. We wish to investigate these proceses (nuclear binding and transport) in oocytes, using nucleoplasmin as a model. Nucleoplasmin is a large endogenous oocyte protein (165,000 daltons) that has a nucleus to cytoplasmic concentration ratio of over 400. It is also known to be transported through the pores. Studies are proposed to establish the location of the transport and binding domains that are present on nucleoplasmin and determine if similar functional sites are associated with other oocytes nuclear proteins. In addition, experiments have been designed to identify the nuclear binding site(s) required for the accumulation of nucleoplasmin and, perhaps, other karyophilic polypeptides. Finally, we hope to determine if the transport process is reversible. These experiments are centered around in vivo procedures and involve the microinjection of affinity purified antibodies against nucleoplasmin and binding proteins.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD020678-02
Application #
3319035
Study Section
Molecular Cytology Study Section (CTY)
Project Start
1985-07-01
Project End
1988-06-30
Budget Start
1986-07-01
Budget End
1987-06-30
Support Year
2
Fiscal Year
1986
Total Cost
Indirect Cost
Name
University of Florida
Department
Type
Schools of Medicine
DUNS #
073130411
City
Gainesville
State
FL
Country
United States
Zip Code
32611
Feldherr, C M; Paine, P L; Hodges, P (1988) Nuclear protein synthesis in animal and vegetal hemispheres of Xenopus oocytes. Exp Cell Res 179:527-34
Dworetzky, S I; Lanford, R E; Feldherr, C M (1988) The effects of variations in the number and sequence of targeting signals on nuclear uptake. J Cell Biol 107:1279-87
Dworetzky, S I; Feldherr, C M (1988) Translocation of RNA-coated gold particles through the nuclear pores of oocytes. J Cell Biol 106:575-84