TAILIEUCHUNG - Báo cáo khoa học: Molecular and functional characterization of novel CRFR1 isoforms from the skin

In our continued studies on corticotropin releasing factor receptor (CRFR1) signaling in the skin, we tested functional activity of CRFR1a, e, f, g and h isoforms after transfection to COS membrane-bound and soluble variants are translatedin vivointofinal proteinproducts that undergo further post-translational the only isoform coupled directly to adenylate cyclase with the exception of an artificial isoform (CRFR1h2) with the insertion of 37 amino acids between the ligand binding domain and the first extracellular loop that was capable of producing detectable levels of cyclic AMP (cAMP) | Eur. J. Biochem. 271 2821-2830 2004 FEBS 2004 doi Molecular and functional characterization of novel CRFR1 isoforms from the skin Alexander Pisarchik and Andrzej Slominski Department of Pathology and Laboratory Medicine University of Tennessee Health Science Center Memphis TN USA In our continued studies on corticotropin releasing factor receptor CRFR1 signaling in the skin we tested functional activity of CRFR1a e f g and h isoforms after transfection to COS cells. t ohi membrane-bound and soluble variants are translated in vivo into final protein products that undergo further post-translational modifications. CRFR1 a was the only isoform coupled directly to adenylate cyclase with the exception of an artificial isoform CRFR1h2 with the insertion of 37 amino acids between the ligand binding domain and the first extracellular loop that was capa-le of producing detectable levels of cyclic AMP cAMP . SohrHe isoforms could modulate cell response with CRFR1e attenuating and CRFR1h amplifying CRFR1a-coupled cAMP production stimulated by urocortin. Testing with plasmids containing the luciferase reporter gene and inducible cis-elements CRE CaRE SRE AP1 or NF-jB demonstrated that only CRFR1a was involved directly in the transcriptional regulation while CRFR1g inhibited CRE higher reporter gene expression by CRF was observed than that mediated by 4b-phorbol 12- myristate 13-acetate and forskolin alone being compatible with the concomitant treatment by phorbol 12-myristate 13- acetate and forskolin. This lnggetSs fiiat bofii pirn tern kinase A and C can be involved in CRF-dependent signal transduction. Keywords skin CRFR1 CRF urocortin COS cells. Corticotropin releasing factor receptors CRFRs are recognized as main central regulators in the humoral and behavioral responses to systemic stress 1-4 . They atso play an important role in the regulation of peripheral organ functions 2 3 5 6 . In the kirn h y may serve as .

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