Most eucaryotic genes are subject to multifactorial regulation by signaling pathways that are triggered by developmental, hormonal, tissue- specific, and/or environmental cues. These pathways often activate complex transcriptional cascades that give rise to altered amounts of protein encoded by target genes hours and even days after the initial signal. Our long term goal is to understand how these complex regulatory events are coordinated to elicit appropriate levels of gene expression. The overall aim of this proposal is to investigate the transcriptional cascades that are evoked by the binding of steroid hormones to their cognate receptors. In particular, our goal is to investigate how estrogen and glucocorticoid induce """"""""secondary"""""""" or late response genes. The ovalbumin gene is indirectly induced by estrogen, glucocorticoid, androgen, and progesterone. Thus, this gene is an excellent model system for investigating the molecular events linking the binding of steroid receptors to early response genes with the subsequent activation of late response genes.
The Specific Aims of this grant are to: I. Ascertain how estrogen and corticosterone induce expression of the ovalbumin gene; and II. Define the role of the negative regulatory element in the positive and negative regulation of the ovalbumin gene. To address the first aim, the plan is to clone two steroid-responsive, cycloheximide-sensitive proteins by using the yeast one-hybrid system. In vivo footprinting techniques will provide the basis for investigating several important questions. These include determining how estrogen and corticosterone synergize to induce transcription of the ovalbumin gene and ascertaining whether single-stranded DNA binding proteins are involved in transcriptional regulation. Mutagenesis and transfection experiments will be employed to continue the functional analysis of the NRE. As steroid hormones have been implicated in the etiology of some cancers, a better. understanding of the molecular events provoked by steroids can only improve the treatment and prevention of these malignancies. Furthermore, such knowledge may provide insights into the development of better contraceptives and aid in the treatment of reproductive and metabolic disorders.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK040082-12
Application #
2905375
Study Section
Biochemical Endocrinology Study Section (BCE)
Program Officer
Margolis, Ronald N
Project Start
1988-05-01
Project End
2000-06-30
Budget Start
1999-07-15
Budget End
2000-06-30
Support Year
12
Fiscal Year
1999
Total Cost
Indirect Cost
Name
University of Minnesota Twin Cities
Department
Biochemistry
Type
Schools of Medicine
DUNS #
168559177
City
Minneapolis
State
MN
Country
United States
Zip Code
55455
Hurt, Elaine M; Saykally, Jessica N; Anose, Bynthia M et al. (2008) Expression of the ZEB1 (deltaEF1) transcription factor in human: additional insights. Mol Cell Biochem 318:89-99
Dougherty, Dawne C; Sanders, Michel M (2005) Comparison of the responsiveness of the pGL3 and pGL4 luciferase reporter vectors to steroid hormones. Biotechniques 39:203-7
Dougherty, Dawne C; Sanders, Michel M (2005) Estrogen action: revitalization of the chick oviduct model. Trends Endocrinol Metab 16:414-9
Dillner, Naomi B; Sanders, Michel M (2004) Transcriptional activation by the zinc-finger homeodomain protein delta EF1 in estrogen signaling cascades. DNA Cell Biol 23:25-34
Monroe, David G; Berger, Ryan R; Sanders, Michel M (2002) Tissue-protective effects of estrogen involve regulation of caspase gene expression. Mol Endocrinol 16:1322-31
Dillner, Naomi B; Sanders, Michel M (2002) Upstream stimulatory factor (USF) is recruited into a steroid hormone-triggered regulatory circuit by the estrogen-inducible transcription factor delta EF1. J Biol Chem 277:33890-4
Berger, R R; Sanders, M M (2000) Estrogen modulates HNF-3beta mRNA levels in the developing chick oviduct. DNA Cell Biol 19:103-12
Monroe, D G; Jin, D F; Sanders, M M (2000) Estrogen opposes the apoptotic effects of bone morphogenetic protein 7 on tissue remodeling. Mol Cell Biol 20:4626-34
Sensenbaugh, K R; Sanders, M M (1999) Multiple promoter elements including a novel repressor site modulate expression of the chick ovalbumin gene. DNA Cell Biol 18:147-56
Chamberlain, E M; Sanders, M M (1999) Identification of the novel player deltaEF1 in estrogen transcriptional cascades. Mol Cell Biol 19:3600-6

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