Aldosterone and vasopressin affect {alpha}- and {gamma}-ENaC mRNA translation.
Vasopressin and aldosterone play key roles in the fine adjustment of sodium and water re-absorption in the nephron. The molecular target of this regulation is the epithelial sodium channel (ENaC) consisting of α-, β- and γ-subunits. We investigated mRNA-specific post-transcriptional mechanisms in hormone-dependent expression of ENaC subunits in mouse kidney cortical collecting duct cells. Transcription experiments and polysome gradient analysis demonstrate that both hormones act on transcription and translation. RNA-binding proteins (RBPs) and mRNA sequence motifs involved in translational control of γ-ENaC synthesis were studied. γ-ENaC-mRNA 3'-UTR contains an AU-rich element (ARE), which was shown by RNA affinity chromatography to interact with AU-rich element binding proteins (ARE-BP) like HuR, AUF1 and TTP. Some RBPs co-localized with γ-ENaC mRNA in polysomes in a hormone-dependent manner. Reporter gene co-expression experiments with luciferase γ-ENaC 3'-UTR constructs and ARE-BP expression plasmids demonstrate the importance of RNA-protein interaction for the up-regulation of γ-ENaC synthesis. We document that aldosterone and the V(2) receptor agonist dDAVP act on synthesis of α- and γ-ENaC subunits mediated by RBPs as effectors of translation but not by mRNA stabilization. Immunoprecipitation and UV-crosslinking analysis of γ-ENaC-mRNA/HuR complexes document the significance of γ-ENaC-mRNA-3'-UTR/HuR interaction for hormonal control of ENaC synthesis.
Perlewitz
Andrea
A
Institut für Vegetative Physiologie, Universitätsmedizin Berlin (Charité), D-10117 Berlin, Germany.
Nafz
Benno
B
Skalweit
Angela
A
Fähling
Michael
M
Persson
Pontus B
PB
Thiele
Bernd-Joachim
BJ
D020413:3' Untranslated Regions; D000450:Aldosterone / Q000494:pharmacology*; D000818:Animals; D000954:Antigens, Surface / Q000378:metabolism; D002478:Cells, Cultured; D003894:Deamino Arginine Vasopressin / Q000494:pharmacology*; D051959:ELAV Proteins; D000067780:ELAV-Like Protein 1; D053503:Epithelial Sodium Channels / Q000096:biosynthesis / Q000235:genetics*; D017930:Genes, Reporter; D007685:Kidney Tubules, Collecting / Q000378:metabolism; D051379:Mice; D011132:Polyribosomes / Q000737:chemistry; D014176:Protein Biosynthesis / Q000187:drug effects; D021122:Protein Subunits / Q000096:biosynthesis / Q000235:genetics; D020871:RNA Stability / Q000187:drug effects; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000032:analysis / Q000378:metabolism; D012261:Ribonucleoproteins / Q000737:chemistry; D015854:Up-Regulation
D020413:3' Untranslated Regions; D000954:Antigens, Surface; D051959:ELAV Proteins; D000067780:ELAV-Like Protein 1; C098770:ELAVL1 protein, human; D053503:Epithelial Sodium Channels; D021122:Protein Subunits; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D012261:Ribonucleoproteins; C026452:messenger ribonucleoprotein; D000450:Aldosterone; D003894:Deamino Arginine Vasopressin
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