The actions of progesterone, a hormone of central importance in controlling the function and growth of female reproductive tissues, such as the breast and uterus, appear to be mediated via interaction with an intracellular protein, the progesterone receptor (PR). Although much is known about the nature of this protein, little is known about the synthesis and degradation of PR and factors that regulate these rates. Our initial studies have provided evidence for a biosynthetic, non hormone-binding precursor of PR and for differences in the form of PR when it is associated with the antiprogestin RU486 vs progestin.
Our aims are to study hormonal factors involved in regulation of PR levels and turnover, to investigate biosynthetic precursors of PR, and to examine the nature of nuclear PR complexes. We will use the density shift technique to examine how turnover kinetics are affected by the nature of the ligand (progestin vs antiprogestin) and the level of receptor occupancy. We will compare systems in which PR is under estrogen control (MCF-7 human breast cancer and rat uterine cells) and independent of estrogen (T47D human brease cancer cells). We will utilize a kinetic model, from which the PR precursor pool size and biosynthetic, activation and degradation rate constants can be derived, and we will attempt to characterize the precursor using monoclonal antibodies to PR. These approaches will also enable us to address the important issue of whether estradiol increases PR levels by changes in receptor synthesis, precursor activation, or stabilization of PR. Since we have found that nuclear PR complexes with antiprogestin sediment as 6 S species under conditions where the complexes with progestin are exclusively 4 S, we will examine factors that affect the 6 S/4 S ratio, determine the subunit composition of the 6 S species by crosslinking and photoaffinity labelling, and evaluate whether the 6 S species represents an """"""""unactivated"""""""" non-DNA binding form of PR. These studies should provide new information on the biosynthesis and degradation of this important regulatory protein, and on the receptor interactions that may underlie progestin antagonist action.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD021524-03
Application #
3320423
Study Section
Biochemical Endocrinology Study Section (BCE)
Project Start
1986-04-01
Project End
1989-11-30
Budget Start
1988-04-01
Budget End
1989-11-30
Support Year
3
Fiscal Year
1988
Total Cost
Indirect Cost
Name
University of Illinois Urbana-Champaign
Department
Type
Schools of Arts and Sciences
DUNS #
041544081
City
Champaign
State
IL
Country
United States
Zip Code
61820
Katzenellenbogen, B S; Nardulli, A M; Read, L D (1990) Estrogen regulation of proliferation and hormonal modulation of estrogen and progesterone receptor biosynthesis and degradation in target cells. Prog Clin Biol Res 322:201-11
Katzenellenbogen, B S; Norman, M J (1990) Multihormonal regulation of the progesterone receptor in MCF-7 human breast cancer cells: interrelationships among insulin/insulin-like growth factor-I, serum, and estrogen. Endocrinology 126:891-8
Read, L D; Greene, G L; Katzenellenbogen, B S (1989) Regulation of estrogen receptor messenger ribonucleic acid and protein levels in human breast cancer cell lines by sex steroid hormones, their antagonists, and growth factors. Mol Endocrinol 3:295-304
Nardulli, A M; Katzenellenbogen, B S (1988) Progesterone receptor regulation in T47D human breast cancer cells: analysis by density labeling of progesterone receptor synthesis and degradation and their modulation by progestin. Endocrinology 122:1532-40
Nardulli, A M; Greene, G L; O'Malley, B W et al. (1988) Regulation of progesterone receptor messenger ribonucleic acid and protein levels in MCF-7 cells by estradiol: analysis of estrogen's effect on progesterone receptor synthesis and degradation. Endocrinology 122:935-44
Read, L D; Snider, C E; Miller, J S et al. (1988) Ligand-modulated regulation of progesterone receptor messenger ribonucleic acid and protein in human breast cancer cell lines. Mol Endocrinol 2:263-71
Katzenellenbogen, B S; Kendra, K L; Norman, M J et al. (1987) Proliferation, hormonal responsiveness, and estrogen receptor content of MCF-7 human breast cancer cells grown in the short-term and long-term absence of estrogens. Cancer Res 47:4355-60
Mullick, A; Katzenellenbogen, B S (1986) Progesterone receptor synthesis and degradation in MCF-7 human breast cancer cells as studied by dense amino acid incorporation. Evidence for a non-hormone binding receptor precursor. J Biol Chem 261:13236-46