Vitamin A is needed by the body to regulate expression of numerous genes with diverse cellular functions. The effects of vitamin A on gene expression arise through interactions of all-trans or 9-cis retinoic acid with members of two families of ligand dependent transcription factors, the retinoic acid receptors (RARs) and the retinoid X receptors (RXRs). All-trans retinoic acid binds only to RARs with high affinity; whereas its 9-cis isomer binds with high affinity to both RARs and RXRs. The actions of all-trans and 9-cis retinoic acid in regulating cellular responses are distinct and not interchangeable. Only recently has convincing data become available regarding the enzymatic formation of all-trans retinoic acid from its precursor retinol and there is still a near complete absence of data regarding how 9-cis retinoic acid is formed within tissues. We have obtained and characterized a full length cDNA clone which, when expressed in CHO cells, avidly catalyzes oxidation of 9-cis retinol to 9-cis retinaldehyde; the first enzymatic step needed for the formation of 9- cis retinoic acid. The CHO cell expressed cDNA does not catalyze all- trans retinol oxidation. Based on this result and other data described in this application, it is clear that we have identified a novel steriospecific enzyme, 9-cis retinol dehydrogenase (9cRDH) which may play an important role in 9-cis retinoic acid formation. This proposal consists of 4 Aims which comprehensively explore the biochemistry and physiology of this novel enzyme.
In Aim 1, we will define the biochemical properties of human and murine 9cRDH and generate antibodies to recombinant forms of the proteins. We will employ, in Aim 2, in situ hybridization and immunocytochemical techniques to explore the cellular localization of 9cRDH in adult mouse tissues and its spatial and temporal pattern of distribution in mouse embryos. Using the highly characterized mouse testis system as a model, in Aim 3, relationships between 9cRDH expression pattern and the actions of vitamin A in maintaining normal testis function will be identified.
Aim 4 will investigate the physiologic significance of the 9cRDH using the techniques of targeted gene disruption.
| Paik, Jisun; Blaner, William S; Swisshelm, Karen (2005) Cis-retinol dehydrogenase: 9-cis-retinol metabolism and its effect on proliferation of human MCF7 breast cancer cells. Exp Cell Res 303:183-96 |
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| Baron, Jens M; Heise, Ruth; Blaner, William S et al. (2005) Retinoic acid and its 4-oxo metabolites are functionally active in human skin cells in vitro. J Invest Dermatol 125:143-53 |
| Piantedosi, Roseann; Ghyselinck, Norbert; Blaner, William S et al. (2005) Cellular retinol-binding protein type III is needed for retinoid incorporation into milk. J Biol Chem 280:24286-92 |
| Paik, Jisun; Vogel, Silke; Quadro, Loredana et al. (2004) Vitamin A: overlapping delivery pathways to tissues from the circulation. J Nutr 134:276S-280S |
| Quadro, Loredana; Blaner, William S; Hamberger, Leora et al. (2004) The role of extrahepatic retinol binding protein in the mobilization of retinoid stores. J Lipid Res 45:1975-82 |
| Quadro, Loredana; Hamberger, Leora; Gottesman, Max E et al. (2004) Transplacental delivery of retinoid: the role of retinol-binding protein and lipoprotein retinyl ester. Am J Physiol Endocrinol Metab 286:E844-51 |
| Thompson, Carol L; Selby, Christopher P; Van Gelder, Russell N et al. (2004) Effect of vitamin A depletion on nonvisual phototransduction pathways in cryptochromeless mice. J Biol Rhythms 19:504-17 |
| Wendler, Christopher C; Schmoldt, Angela; Flentke, George R et al. (2003) Increased fibronectin deposition in embryonic hearts of retinol-binding protein-null mice. Circ Res 92:920-8 |
| Paik, Jisun; Blaner, William S; Sommer, Karen M et al. (2003) Retinoids, retinoic acid receptors, and breast cancer. Cancer Invest 21:304-12 |
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