title: Transactivation properties of retinoic acid and retinoid X receptors in mammalian cells and yeast. Correlation with hormone binding and effects of metabolism.
abstract: The binding affinities of 9-cis-retinoic acid (9-cis-RA) and all-trans-retinoic acid (t-RA) for retinoic acid receptors (RAR) alpha, beta, and gamma and for retinoid X receptors (RXR) alpha, beta, and gamma were determined using the recombinant receptor proteins and were compared with each hormone's ability to activate transcription through the receptors in mammalian and yeast cell systems. 9-cis-RA bound to both the RXRs (Kd values = 1.4-2.4 nM) and the RARs (Kd values = 0.2-0.8 nM). The ability of 9-cis-RA to bind to the RARs and RXRs correlated with its ability to produce similar transactivation profiles with these receptors in mammalian and yeast cell assays. t-RA bound to the RARs (Kd values = 0.2-0.4 nM) and activated transcription through the RARs in mammalian and yeast cells. In contrast, while t-RA did not bind to the RXRs, it did activate the RXRs, albeit less potently than 9-cis-RA, in mammalian cells. In yeast, however, the RXRs activated transcription only in the presence of 9-cis-RA, not with t-RA. While RAR gamma is activated in yeast by either t-RA or 9-cis-RA, the overall level of transcription was increased upon the addition of hormone-occupied RXR. Metabolism studies suggest that while there was no cell-dependent interconversion between t-RA and 9-cis-RA in yeast, there was cell-dependent conversion of 9-cis-RA to t-RA in mammalian cells [corrected].
authors.affiliation: Ligand Pharmaceuticals, Inc., Department of Biochemistry, San Diego, California 92121.
authors.forename: E A
authors.initials: EA
authors.lastname: Allegretto
authors.forename: M R
authors.initials: MR
authors.lastname: McClurg
authors.forename: S B
authors.initials: SB
authors.lastname: Lazarchik
authors.forename: D L
authors.initials: DL
authors.lastname: Clemm
authors.forename: S A
authors.initials: SA
authors.lastname: Kerner
authors.forename: M G
authors.initials: MG
authors.lastname: Elgort
authors.forename: M F
authors.initials: MF
authors.lastname: Boehm
authors.forename: S K
authors.initials: SK
authors.lastname: White
authors.forename: J W
authors.initials: JW
authors.lastname: Pike
authors.forename: R A
authors.initials: RA
authors.lastname: Heyman
mesh_terms: D000818:Animals; D001665:Binding Sites; D015153:Blotting, Western; D002460:Cell Line; D005786:Gene Expression Regulation / Q000502:physiology; D006728:Hormones / Q000378:metabolism; D015474:Isotretinoin / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D018168:Receptors, Retinoic Acid / Q000378:metabolism*; D047488:Retinoid X Receptors; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014157:Transcription Factors; D015533:Transcriptional Activation; D014212:Tretinoin / Q000378:metabolism
chemical_list: D006728:Hormones; D018160:Receptors, Cytoplasmic and Nuclear; D018168:Receptors, Retinoic Acid; D047488:Retinoid X Receptors; D014157:Transcription Factors; D014212:Tretinoin; D015474:Isotretinoin