Skip to main content
SpringerLink
Log in

Studies of the type I cellular retinoic acid-binding protein mutants and their biological activities

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. 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

    Google Scholar 

  2. Glass CK: Differential recognition of target genes by nuclear receptor monomers, dimers and heterodimers. Endocrine Rev 15: 391–407, 1994

    Google Scholar 

  3. Sucov HM, Evans RM: Retinoic acid and retinoic acid receptors in development. Mol Neurobiol 10: 169–184,1995

    PubMed  Google Scholar 

  4. 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

    Google Scholar 

  5. Ong DE: Cellular transport and metabolism of vitamin A: Roles of the cellular retinoid-binding proteins. Nutr Rev 52: S24–S31, 1994

    Google Scholar 

  6. Napoli JL: Retinoid binding proteins redirect retinoid metabolism: Biosynthesis and metabolism of retinoic acid. Sem Cell Dev Biol 8: 403–415, 1997

    Google Scholar 

  7. 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

    PubMed  Google Scholar 

  8. 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

    PubMed  Google Scholar 

  9. 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

    Google Scholar 

  10. 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

    PubMed  Google Scholar 

  11. 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

    Google Scholar 

  12. 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

    Google Scholar 

  13. 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

    PubMed  Google Scholar 

  14. 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

    PubMed  Google Scholar 

  15. 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

    Google Scholar 

  16. 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

    Google Scholar 

  17. 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

    PubMed  Google Scholar 

  18. 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

    PubMed  Google Scholar 

  19. 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

    PubMed  Google Scholar 

  20. 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

    PubMed  Google Scholar 

  21. 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

    PubMed  Google Scholar 

  22. Wang L, Yue L, Yan H: Structure finction relationships of cellular retinoic acid binding protein. J Biol Chem 272: 1541–1547, 1997

    PubMed  Google Scholar 

  23. 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

    PubMed  Google Scholar 

  24. 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

    PubMed  Google Scholar 

  25. 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

    PubMed  Google Scholar 

  26. 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

    PubMed  Google Scholar 

  27. 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

    PubMed  Google Scholar 

  28. 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

    Google Scholar 

  29. 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

    Google Scholar 

  30. 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

    PubMed  Google Scholar 

  31. 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

    Google Scholar 

  32. 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

    PubMed  Google Scholar 

  33. 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

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Pharmacology, University of Minnesota Medical School, Minneapolis, MN, 55455, USA

    Li-Na Wei, Liming Chang & Xinli Hu

Authors
  1. Li-Na Wei
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Liming Chang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xinli Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints 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

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006906415388

Search

Navigation