Arginine of retinoic acid receptor beta which coordinates with the carboxyl group of retinoic acid functions independent of the amino acid residues responsible for retinoic acid receptor subtype ligand specificity.
The biological actions of retinoic acid (RA) are mediated by retinoic acid receptors (RARalpha, RARbeta, and RARgamma) and retinoid X receptors (RXRalpha, RXRbeta, and RXRgamma). Consistent with the X-ray crystal structures of RARalpha and RARgamma, site-directed mutagenesis studies have demonstrated the importance of a conserved Arg residue (alphaArg(276), betaArg(269), and gammaArg(278)) for coordination with the carboxyl group of RA. However, mutation of Arg(269) to Ala in RARbeta causes only a 3- to 6-fold increase in the K(d) for RA and EC(50) in RA-dependent transcriptional transactivation assays while the homologous mutation in either RARalpha or RARgamma causes a 110-fold and a 45-fold increase in EC(50) value, respectively. To further investigate the nature of this difference, we prepared mutant RARs to determine the effect of conversion of betaR269A to a mutant which mimics either RARalpha ligand selectivity (betaA225S/R269A) or RARgamma ligand selectivity (betaI263M/R269A/V338A). Our results demonstrate that in RARbeta mutants that acquire either RARalpha or RARgamma ligand specificity the Arg(269) position responsible for coordination with the carboxyl group of retinoids continued to function like that of RARbeta. Furthermore, three mutant receptors (betaA225S/R269A, betaA225S/F279, and alphaF286A) were found to have a greater than wild-type affinity for the RARalpha-selective ligand Am580. Finally, a homology-based computer model of the ligand binding domain (LBD) of RARbeta and the X-ray crystal structures of the LBD of both RARalpha and RARgamma are used to describe potential mechanisms responsible for the increased affinity of some mutants for Am580 and for the difference in the effect of mutation of Arg(269) in RARbeta compared to its homologous Arg in RARalpha and RARgamma.
Zhang
Zeng Ping
ZP
Department of Biochemistry, Temple University School of Medicine, 3420 N. Broad St., Philadelphia, PA 19140, USA.
Hutcheson
Juliet M
JM
Poynton
Helen C
HC
Gabriel
Jerome L
JL
Soprano
Kenneth J
KJ
Soprano
Dianne Robert
DR
D000409:Alanine / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019943:Amino Acid Substitution; D000596:Amino Acids / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001120:Arginine / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D001565:Benzoates / Q000819:agonists / Q000378:metabolism; D002245:Carbon Dioxide / Q000737:chemistry*; D008024:Ligands; D051379:Mice; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D018168:Receptors, Retinoic Acid / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D012176:Retinoids / Q000819:agonists / Q000378:metabolism; D012680:Sensitivity and Specificity; D013329:Structure-Activity Relationship; D013764:Tetrahydronaphthalenes / Q000819:agonists / Q000378:metabolism; D014212:Tretinoin / Q000737:chemistry* / Q000378:metabolism*; D000097768:Retinoic Acid Receptor gamma
D000409:Alanine; D000596:Amino Acids; D001120:Arginine; D001565:Benzoates; D002245:Carbon Dioxide; D008024:Ligands; D018168:Receptors, Retinoic Acid; D011993:Recombinant Fusion Proteins; D012176:Retinoids; D013764:Tetrahydronaphthalenes; D014212:Tretinoin; D000097768:Retinoic Acid Receptor gamma; C099555:CD 437; C082925:retinoic acid receptor beta; C068073:Am 580; C041069:carboxyl radical