Post-transcriptional Regulation of Soluble Guanylyl Cyclase Expression in Rat Aorta
| Autor: | Alexander Mülsch, Stephan Klöß, Henry Furneaux |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Male
Untranslated region Time Factors Transcription Genetic Receptors Cytoplasmic and Nuclear Biochemistry ELAV-Like Protein 1 Soluble Guanylyl Cyclase RNA Processing Post-Transcriptional RNA Small Interfering 3' Untranslated Regions Aorta Cells Cultured Oxadiazoles Reverse Transcriptase Polymerase Chain Reaction RNA-Binding Proteins Guanylate cyclase 2C ELAV Proteins Antigens Surface Dactinomycin Protein Binding Indazoles Protein subunit Blotting Western Molecular Sequence Data Down-Regulation Enzyme Activators Alpha (ethology) Biology Transfection Enzyme activator Oxazines Animals RNA Messenger Rats Wistar Molecular Biology Nucleic Acid Synthesis Inhibitors Cell Nucleus Messenger RNA Base Sequence Three prime untranslated region Muscle Smooth Cell Biology Blotting Northern Molecular biology Protein Structure Tertiary Rats Guanylate Cyclase RNA Poly A Soluble guanylyl cyclase |
| Zdroj: | Journal of Biological Chemistry. 278:2377-2383 |
| ISSN: | 0021-9258 |
| Popis: | We investigated the molecular mechanism of cyclic GMP-induced down-regulation of soluble guanylyl cyclase expression in rat aorta. 3-(5'-Hydroxymethyl-2'-furyl)-1-benzyl indazole (YC-1), an allosteric activator of this enzyme, decreased the expression of soluble guanylyl cyclase alpha(1) subunit mRNA and protein. This effect was blocked by the enzyme inhibitor 4H-8-bromo-1,2,4-oxadiazolo(3,4-d)benz(b-1,4)oxazin-1-one (NS2028) and by actinomycin D. Guanylyl cyclase alpha(1) mRNA-degrading activity was increased in protein extracts from YC-1-exposed aorta and was attenuated by pretreatment with actinomycin D and NS2028. Gelshift and supershift analyses using an adenylate-uridylate-rich ribonucleotide from the 3'-untranslated region of the alpha(1) mRNA and a monoclonal antibody directed against the mRNA-stabilizing protein HuR revealed HuR mRNA binding activity in aortic extracts, which was absent in extracts from YC-1-stimulated aortas. YC-1 decreased the expression of HuR, and this decrease was prevented by NS2028. Similarly, down-regulation of HuR by RNA interference in cultured rat aortic smooth muscle cells decreased alpha(1) mRNA and protein expression. We conclude that HuR protects the guanylyl cyclase alpha(1) mRNA by binding to the 3'-untranslated region. Activation of guanylyl cyclase decreases HuR expression, inducing a rapid degradation of guanylyl cyclase alpha(1) mRNA and lowering alpha(1) subunit expression as a negative feedback response. |
| Databáze: | OpenAIRE |
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