The goal of this proposal is to establish a molecular basis for pattern formation during early vertebrate development. The discovery of the retinoic acid receptor and its homology to the steroid/thyroid receptor superfamily, offers for the first time in a vertebrate system, the hope of analyzing the mechanisms by which positional information are established and thus, how the fate and behavior of embryonic cells are controlled. Utilizing the cloned retinoic acid receptor gene products as models, we will define, in molecular terms, how retinoids influence development and physiology by controlling patterns of gene expression. We will characterize the molecular interactions of the retinoic acid receptor with DNA by identifying putative response elements (RAREs) present in the retinoic acid receptor and osteocalcin genes. In addition to biochemical dissection of the DNA-protein complex, the DNA-binding domain of the RAR and the ligand-binding domain will be overproduced in E. coli to begin NMR and X-ray crystallographic studies. Activation domains of the retinoid receptors will be identified by deletion, truncation, and point mutation analysis. Proteins interacting with the RARs will be sought by chemical approaches to identify the encoding genes. These approaches include chromatography of nuclear extracts to RAR affinity columns and the detection of receptor-DNA complexes with altered electrophoretic mobilities. In situ hybridization and immunohistochemical studies will be used to localize subtype-specific RAR mRNA and protein during mouse embryogenesis. Functional roles of the RARs during early embryonic development will be assessed by creating transgenic animals expressing a variety of mutant RAR derivatives. RAR responsive promoters linked to beta-galactosidase will be used to visualize endogenous RA gradients. In summary, retinoic acid, because of its described role in limb development and its capacity to induce differentiation in embryo carcinoma cells, is an ideal candidate for a vertebrate morphogen. In combination with the recently cloned retinoic acid receptor gene products in several species, we are now poised to examine the mechanism by which RA and the RARs specify pattern formation in vertebrates.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD027183-02
Application #
3328754
Study Section
Molecular Cytology Study Section (CTY)
Project Start
1990-09-12
Project End
1995-08-31
Budget Start
1991-09-01
Budget End
1992-08-31
Support Year
2
Fiscal Year
1991
Total Cost
Indirect Cost
Name
Salk Institute for Biological Studies
Department
Type
DUNS #
005436803
City
La Jolla
State
CA
Country
United States
Zip Code
92037
Uhlenhaut, N Henriette; Barish, Grant D; Yu, Ruth T et al. (2013) Insights into negative regulation by the glucocorticoid receptor from genome-wide profiling of inflammatory cistromes. Mol Cell 49:158-71
Barish, Grant D; Yu, Ruth T; Karunasiri, Malith S et al. (2012) The Bcl6-SMRT/NCoR cistrome represses inflammation to attenuate atherosclerosis. Cell Metab 15:554-62
Murphy, Samantha H; Suzuki, Kotaro; Downes, Michael et al. (2011) Tumor suppressor protein (p)53, is a regulator of NF-kappaB repression by the glucocorticoid receptor. Proc Natl Acad Sci U S A 108:17117-22
Narkar, Vihang A; Fan, Weiwei; Downes, Michael et al. (2011) Exercise and PGC-1?-independent synchronization of type I muscle metabolism and vasculature by ERR?. Cell Metab 13:283-93
Pei, Liming; Leblanc, Mathias; Barish, Grant et al. (2011) Thyroid hormone receptor repression is linked to type I pneumocyte-associated respiratory distress syndrome. Nat Med 17:1466-72
Mihaylova, Maria M; Vasquez, Debbie S; Ravnskjaer, Kim et al. (2011) Class IIa histone deacetylases are hormone-activated regulators of FOXO and mammalian glucose homeostasis. Cell 145:607-21
Yamamoto, Hiroyasu; Williams, Evan G; Mouchiroud, Laurent et al. (2011) NCoR1 is a conserved physiological modulator of muscle mass and oxidative function. Cell 147:827-39
Kacevska, Marina; Downes, Michael R; Sharma, Rohini et al. (2011) Extrahepatic cancer suppresses nuclear receptor-regulated drug metabolism. Clin Cancer Res 17:3170-80
Sugii, Shigeki; Kida, Yasuyuki; Berggren, W Travis et al. (2011) Feeder-dependent and feeder-independent iPS cell derivation from human and mouse adipose stem cells. Nat Protoc 6:346-58
Fang, Sungsoon; Suh, Jae Myoung; Atkins, Annette R et al. (2011) Corepressor SMRT promotes oxidative phosphorylation in adipose tissue and protects against diet-induced obesity and insulin resistance. Proc Natl Acad Sci U S A 108:3412-7

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