David J. Shapiro

Professor of Biochemistry
Professor of Basic Medical Sciences, College of Medicine


B.S. 1967 Brooklyn College
Ph.D. 1972 Purdue University
Postdoc. 1972-1973 Stanford University Medical School, 1973-1974 Stanford University


Office:

Phone:

Fax:

Email:

413 Roger Adams Lab.
217-333-1788
217-244-5858

djshapir@uiuc.edu

Mailing Address:

Department of Biochemistry
419 Roger Adams Lab.
University of Illinois
600 S. Mathews Avenue
Urbana, IL 61801
Lab Website

Estrogen action: from gene transcription and mRNA metabolism to inflammation, apoptosis and immune surveillance

The effects of steroid hormones such as estrogens, testosterone, and thyroid hormone are mediated through specific proteins called steroid/nuclear receptors. Our research centers primarily on the molecular mechanisms by which estrogen receptor (ER) carries out its diverse biological activities. Estrogens, acting via the ER, regulate the growth and differentiation of cells of the female and male reproductive systems, the growth and metatastic potential of breast and uterine cancer cells and have important effects on brain development, liver pathology autoimmune diseases and inflammatory processes, bone metabolism and cardiovascular health. We are studying the mechanisms by which estrogens, acting through ERs, act to control gene transcription and mRNA metabolim and the consequences of those actions in cell biology and human disease.

Tamoxifen is widely used in breast cancer chemotherapy and chemo-prevention. Tamoxifen both competes with estrogens for binding to the estrogen receptor and has the ability to induce apoptosis (programmed cell death) of target breast and ovarian cancer cells. We find that high concentrations of tamoxifen kill cells through an ER-independent pathway while low concentrations of tamoxifen can kill some cancer cells through an ER-dependent pathway. Tamoxifen bound to the ER must activate specific signal transduction pathways and must bind to DNA to induce death of cancer cells. These studies have led us to a new model for how estrogen bound to ER regulates transcription of specific genes.

We recently described proteinase inhibitor 9 (PI-9), a novel estrogen regulated gene in humans, which is likely to play an important role in the well-known ability of estrogens to protect against several inflammatory and immune diseases. PI-9 inhibits granzyme B, the primary protease immune system cells used to kill off transformed or infected cells. In collaboration with the laboratory of Prof. D. Kranz we recently showed that estrogen induction of PI-9 protects liver cells against apoptosis induced by cytotoxic T lymphocytes (CTLs). This is the first example of estrogen regulating the ability of a target cell to be killed off by the immune surveillance system. Our finding is likely related to the protective effects of estrogen against liver cancer.

To investigate hormone control of mRNA stability, we developed a model system based on our finding that estrogen induces a 30 fold increase in the cytoplasmic stability of vitellogenin mRNA. We identified vigilin, as an estrogen-inducible protein, which binds to a segment of the mRNA important in estrogen?mediated mRNA stabilization. Remarkably, vigilin contains 15 RNA binding domains. We used RNA interference to show that vigilin is essential for the viability of human cells. RNAi knockdown of vigilin triggers rapid apoptosis of human cells. In collaborative studies (with the labs of Profs. D. Schoenberg and G. Carmichael) we are analyzing the roles of vigilin in the regulation of mRNA stability and in nuclear RNA editing.

For background and more details see our Web site at www.life.uiuc.edu/shapiro/. This site is also accessible through this entry in the Dept. of Biochemistry Web site.


Representative Publications:

Mechanism of Estrogen Receptor Regualtion of Gene Transcription
Krieg, A.J., Krieg, S.A., Ahn, B.S., and Shapiro, D.J. (2004) "Interplay Between Estrogen Response Element Sequence and Ligands Controls in Vivo Binding of Estrogen Receptor to Regulated Genes," J. Biol. Chem. 279, 5025-34. [Abstract]

Wang, S., Ahn, B.S., Harris, R., Nodeen, S.K. and Shapiro, D.J. (2004) "A Fluorescence Anisotropy Microplate Assay for Analysis of Steroid Receptor DNA Interactions," Submitted for publication. [Abstract]

Tamoxifen-induced Apoptosis
Obrero, M., Yu, D.V., and Shapiro, D.J. (2002) "Estrogen Receptor-dependent and Estrogen Receptor-independent Pathways for Tamoxifen and 4-hydroxytamoxifen-induced Programmed Cell Death," J. Biol. Chem. 277, 45695-703. [Abstract]

Regulation and Function of Proteinase Inhibitor 9 (PI-9)
Kannan-Thulasiraman, P. and Shapiro, D.J. (2002) "Modulators of Inflammation Use Nuclear Factor-kappa B and Activator Protein-1 Sites to Induce the Caspase-1 and Granzyme B Inhibitor, Proteinase Inhibitor 9," J. Biol. Chem. 277, 41230-9. [Abstract]

Estrogen and Vigilin Control of RNA Metabolism
Dodson, R.E., Goolsby, K.M., Acena-Nagel. M., Mao, C. and Shapiro, D.J. (2003) "RNA Gel Shift Assays for Analysis of Hormone Control of mRNA Stability," Methods in Enzymology 364, 350-61. [Abstract]

Goolsby, K.M. and Shapiro, D.J. (2003) "RNAi-mediated Depletion of the 15 KH Domain Protein, Vigilin, Induces Death of Dividing and Non-dividing Human Cells but does not Initially Inhibit Protein Synthesis," Nucleic Acids Res. 31, 5644-53. [Abstract]


Publications by David J. Shapiro listed on the National Library of Medicine (PubMed)