Abstract
We have mutated the type I cellular retinoic acid binding protein (CRABP-I), individually at the Arg131 (into Ala) and the Tyr 133 (into Phe) residues which have been predicted to make direct contact with retinoic acid (RA) based upon previous structural studies. The RA-binding affinities of these mutants are examined and their biological effects on RA induction of reporter genes are determined. The R131A mutation drastically affects its ligand-binding property, but the Y133F mutation has little effect. By using an RA-inducible reporter, it is found that the wild type CRABP-I exerts biphasic effects on RA induction of the reporter. The early (at 12 h) effect is to enhance RA induction, whereas the delayed (at 24 h) effect is to suppress RA induction. In consistence with their RA binding property, the R 131A mutant loses both its early and delayed biological activities, whereas the Y133F mutant remains as effective as the wild type. It is concluded that CRABP-I over-expression exerts biphasic effects on RA-mediated gene expression, and that Arg131, but not Tyr 133, is essential for a high RA-binding affinity of this protein as well as its biological activity.
Similar content being viewed by others
References
Mangelsdorf DJ, Umesono, Evans RM: The retinoid receptors. In: M.B. Sporn, A.B. Roberts, D.S. Goodman (eds). The Retinoids, Vol. 2. Academic Press, New York, 1993, pp 319–350
Glass CK: Differential recognition of target genes by nuclear receptor monomers, dimers and heterodimers. Endocrine Rev 15: 391–407, 1994
Sucov HM, Evans RM: Retinoic acid and retinoic acid receptors in development. Mol Neurobiol 10: 169–184,1995
Blaner WS, Olson JA: Retinol and retinoic acid metabolism. In: M.B. Sporn, A.B. Roberts, D.S. Goodman (eds). The Retinoids, Vol. 2. Academic Press, New York, 1993, pp 229–255
Ong DE: Cellular transport and metabolism of vitamin A: Roles of the cellular retinoid-binding proteins. Nutr Rev 52: S24–S31, 1994
Napoli JL: Retinoid binding proteins redirect retinoid metabolism: Biosynthesis and metabolism of retinoic acid. Sem Cell Dev Biol 8: 403–415, 1997
Fawcett D, Pascer P, Fraser R, Colbert M, Rossant J, Giguere V: Postaxial polydactylyl in forelimbs of CRABPII mutant mice. Development 121: 671–679, 1995
Lampron C, Rochette-Egly Q, Gorry P, Dolle P, Mark M, Lufkin T, LeMeur M, Chambon P: Mice deficient in cellular retinoic acid binding protein II (CRABPII) or in both CRABP I and CRABP II are essentially normal. Development 121: 539–548, 1995
Gorry P, Lufkin T, Dierich A, Rochette-Egly C, Decimo D, Dolle P, Mark M, Durano B, Chambon P: The cellular retinoic acid binding protein I is dispensable. Proc Natl Sci Acad USA 91: 9032–9036, 1994
de Bruijn DRH, Oerlemans F, Hendriks W, Baats E, Ploemacher R, Wieringa B, van Kessel AG: Normal development, growth and reproduction in cellular retinoic acid binding protein I null mutant mice. Differentiation 58: 141–148, 1994
Wei L-N, Lee C-H, Chang S, Chu Y-S: Pathogenesis in transgenic mice expressing bovine cellular retinoic acid-binding protein. Dev Growth Diff 34: 479–488, 1992
Wei L-N, Chang L, Lee C-H: Studies of over-expressing cellular retinoic acid binding protein I in cultured cells and transgenic mice. Transgenics 2: 201–209, 1997
Gustafson A-L, Donovan M, Annerwall E, Dencker L, Eriksson U: Nuclear import of cellular retinoic acid binding protein type 1in mouse embryonic cells. Mech Dev 58: 27–38, 1996
Venepally P, Reddy LG, Sani BP: Analysis of the effects of CRABP I expression on the RA-induced transcription mediated by retinoid receptors. Biochemistry 35: 9974–9982, 1996
Boylan JF, Gudas U: Overexpression of the cellular retinoic acid binding protein I results in a reduction in differentiation specific gene expression in F9 teratocarcinoma cells. J Cell Biol 112: 1965–1979, 1991
Nugent P, Greene R: Antisense oligonucleotides to CRABP-I and II alter the expression of TGF-β, RAR-β and tenascin in primary cultures of embryonic palate cells. In Vitro Cell Dev Biol 31: 553–558, 1995
el Akawi Z, Napoli JL: Rat liver cytosolic retinal dehydrogenase: Comparison of 13-cis-, 9-cis-, and all-trans-retinal as substrates and effects of cellular retinoid-binding proteins and retinoic acid on activity. Biochemistry 33: 1938–43, 1994
Raner GM, Vaz ADN, Coon MJ: Metabolism of all-trans, 9-cis, and 13-cis isomers of retinal by purified isozymes of inhibition of retinoid oxidation by citral. Mol Pharmacol 49: 515–522, 1996
Rizo J, Liu Z, Gierasch LM: H and 15N resonance assignments and secondary structure of cellular retinoic acid-binding protein with and without bound ligand. J Biomol NMR 4: 741–760,1994
Kleywegt GJ, Bergfors, T, Senn H, Le Motte P, Gsell B, Shudo K, Jones TA: Crystal structures of cellular retinoic acid binding proteins I and II in complex with all-trans retinoic acid and a synthetic retinoid. Structure 2: 1241–1258, 1994
Thompson JR, Bratt JM, Banaszak L: Crystal structure of cellular retinoic acid binding protein I shows increased access to the binding cavity due to formation of an intermolecular β-sheet. J Mol Biol 252: 433–446, 1995
Wang L, Yue L, Yan H: Structure finction relationships of cellular retinoic acid binding protein. J Biol Chem 272: 1541–1547, 1997
Chen LX, Zhang Z-P, Scafonas A, Cavalli RC, Gabriel JL, Soprano KJ, Soprano DR: Arginine 132 of cellular retinoic acid-binding protein (type II) is important for binding of retinoic acid. J Biol Chem 270: 4518–4525, 1995
de The H, Vivanco-Ruiz Mdel M, Tiollais P, Stunnenberg H, Dejean A: Identification of a retinoic acid responsive element in the retinoic acid receptor beta gene. Nature 343: 177–180, 1990
Chang L, Wei L-N: Characterization of a negative response-DNA element in the upstream region of the cellular retinoic acid-binding protein I gene of the mouse. J Biol Chem 272: 10144–10150, 1997
Wei L-N, Mertz JR, Goodman DS, Nguyen-Huu C: Cellular retinoic acid-and cellular retinol-binding proteins: cDNA cloning, chromosomal assignment and tissue specific expression. Mol Endocrinol 1: 526–534, 1987
Wei L-N, Lee C-H, Filipcik P, Chang L: Regulation of the mouse cellular retinoic acid binding protein I gene by thyroid hormone and retinoids in transgenic mouse embryos. J Endocrinol 155: 35–46, 1997
Ausubel TM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Strulil: Analysis of Proteins (Current Protocols in Molecular Biology). John Wiley & Sons, New York 1993, pp 10.0.1–10.5.5
Fiorella P D, Giguere V, Napoli JL: Expression of cellular retinoic acid-binding protein (type II) in Escherichia coli. Characterization and comparison to cellular retinoic acid-binding protein (type I). J Biol Chem 268: 21543–21552, 1993
Cogan U, Kopelman M, Mokady S, Shinitzky M: Binding affinities of retinol and related compounds to retinol binding proteins. Eur J Biochem 65: 71–78, 1976
White JA, Beckett-Jones B, Kuo Y-D, Dilworth FJ, Bonasoro J, Jones G, Petkovich M: cDNA cloning of human retinoic acid metabolizing enzyme (hP45ORAI) identifies a novel family of cytochromes P450 (CYP26). J Biol Chem 272: 18538–18541, 1996
Fujii H, Sato T, Kaneko S, Gotoh O, Fujii-Kuriyama. Y, Osawa K, Kato S, Hamada H: Metabolic inactivation of retinoic acid by a novel p450 differentially expressed in developing mouse embryos. EMBO J 16: 4163–4173,1997
Ray WJ, Bain G, Yao M, Gottlieb DI: CYP26, a novel mammalian cytochrome p450 induced by retinoic acid and defines a new family. J Biol Chem 272: 18702–18708, 1997
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Wei, LN., Chang, L. & Hu, X. Studies of the type I cellular retinoic acid-binding protein mutants and their biological activities. Mol Cell Biochem 200, 69–76 (1999). https://doi.org/10.1023/A:1006906415388
Issue Date:
DOI: https://doi.org/10.1023/A:1006906415388