HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Tate, B. F. Articles by Grippo, J. F. Search for Related Content PubMed PubMed Citation Articles by Tate, B. F. Articles by Grippo, J. F. Social Bookmarking                      What's this?

Volume 270, Number 35, Issue of September 01, pp. 20258-20263, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.

---

Mutagenesis of the Ligand Binding Domain of the Human Retinoic Acid Receptor Identifies Critical Residues for 9-cis-Retinoic Acid Binding

(Received for publication, March 7, 1995; and in revised form, June 13, 1995)

Bonnie F. Tate,  Joseph F. Grippo

We have recently identified a small region (amino acids 405-419) within the ligand binding domain of a truncated human retinoic acid receptor (419) that is required for binding of 9-cis-retinoic acid (RA), but not all-trans-retinoic acid (t-RA). To probe the structural determinants of this high affinity 9-cis-RA binding site, a series of 419 mutants were prepared whereby an individual alanine residue was substituted for each amino acid within this region. These modified receptors were expressed in mammalian COS-1 cells and assayed for their ability to bind 9-cis-RA as well as t-RA. Only two of the mutants, M406A (mutation of methionine 406 to alanine), and I410A (mutation of isoleucine 410 to alanine) exhibit no detectable binding of 9-cis-RA when analyzed using saturation binding kinetics. Substitution of methionine 406 with the amino acids leucine, isoleucine, and valine yields mutant receptors that exhibit decreased binding for 9-cis-RA as the length or hydrophobicity of the R group decreases. Further substitution of methionine 406 with the small polar amino acid, threonine, results in a loss of detectable 9-cis-RA binding. Since amino acids 405-419 on a human RAR (hRAR) are predicted to form a short amphipathic -helix, modeling of this structure into a helical wheel indicates that these two amino acids, methionine 406 and isoleucine 410, are actually positioned proximal to each other. Data presented here suggest that high affinity 9-cis-RA binding to a hRAR depends on an interaction with the two amino acids methionine 406 and isoleucine 410.

---

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				A. Mouchon, M.-H. Delmotte, P. Formstecher, and P. Lefebvre Allosteric Regulation of the Discriminative Responsiveness of Retinoic Acid Receptor to Natural and Synthetic Ligands by Retinoid X Receptor and DNA Mol. Cell. Biol., April 1, 1999; 19(4): 3073 - 3085. [Abstract] [Full Text] [PDF]

				W. Ding, Y.-P. Li, L. M. Nobile, G. Grills, I. Carrera, E. Paietta, M. S. Tallman, P. H. Wiernik, and R. E. Gallagher Leukemic Cellular Retinoic Acid Resistance and Missense Mutations in the PML-RARalpha Fusion Gene After Relapse of Acute Promyelocytic Leukemia From Treatment With All-trans Retinoic Acid and Intensive Chemotherapy Blood, August 15, 1998; 92(4): 1172 - 1183. [Abstract] [Full Text] [PDF]

				I. G. Schulman, G. Shao, and R. A. Heyman Transactivation by Retinoid X Receptor-Peroxisome Proliferator-Activated Receptor gamma  (PPARgamma ) Heterodimers: Intermolecular Synergy Requires Only the PPARgamma Hormone-Dependent Activation Function Mol. Cell. Biol., June 1, 1998; 18(6): 3483 - 3494. [Abstract] [Full Text]

				L. Benedetti, A. A. Levin, B. M. Scicchitano, F. Grignani, G. Allenby, D. Diverio, F. Lo Coco, G. Avvisati, M. Ruthardt, S. Adamo, et al. Characterization of the Retinoid Binding Properties of the Major Fusion Products Present in Acute Promyelocytic Leukemia Cells Blood, August 1, 1997; 90(3): 1175 - 1185. [Abstract] [Full Text] [PDF]

				S. Keidel, F. P. Y. Lamour, and C. M. Apfel Mutational Analysis Reveals That All-trans-retinoic acid, 9-cis-Retinoic acid, and Antagonist Interact with Distinct Binding Determinants of RARalpha J. Biol. Chem., July 18, 1997; 272(29): 18267 - 18272. [Abstract] [Full Text] [PDF]

				I G Schulman, C Li, J W Schwabe, and R M Evans The phantom ligand effect: allosteric control of transcription by the retinoid X receptor. Genes & Dev., February 1, 1997; 11(3): 299 - 308. [Abstract] [PDF]

				J. E. Driscoll, C. L. Seachord, J. A. Lupisella, R. P. Darveau, and P. R. Reczek Ligand-induced Conformational Changes in the Human Retinoic Acid Receptor gamma Detected Using Monoclonal Antibodies J. Biol. Chem., September 20, 1996; 271(38): 22969 - 22975. [Abstract] [Full Text] [PDF]

				M. A.C. Pratt, D. Deonarine, C. Teixeira, D. Novosad, B. F. Tate, and J. F. Grippo The AF-2 Region of the Retinoic Acid Receptor alpha Mediates Retinoic Acid Inhibition of Estrogen Receptor Function in Breast Cancer Cells J. Biol. Chem., August 23, 1996; 271(34): 20346 - 20352. [Abstract] [Full Text] [PDF]

				C. Bastie, S. Luquet, D. Holst, C. Jehl-Pietri, and P. A. Grimaldi Alterations of Peroxisome Proliferator-activated Receptor delta Activity Affect Fatty Acid-controlled Adipose Differentiation J. Biol. Chem., December 1, 2000; 275(49): 38768 - 38773. [Abstract] [Full Text] [PDF]