Angiotensin II and stretch activate NADPH oxidase to destabilize cardiac Kv4.3 channel mRNA.
Pathological and physiological hypertrophy of the heart is associated with decreased expression of the Kv4.3 transient outward current (Ito) channel. The downregulation of channel mRNA and protein, which may be proarrhythmic, is recapitulated with cultured neonatal rat ventricular myocytes treated with angiotensin II (Ang II). Here we show that the 4.9 kb 3' untranslated region (3' UTR) of the Kv4.3 channel transcript confers Ang II sensitivity to a promoter-reporter construct. In contrast, Kv4.2 and Kv1.5 3'-UTR sequences are insensitive to Ang II. Both Kv4.3 3'-UTR reporter mRNA and activity are decreased in Ang II-treated cardiac myocytes, in accordance with a decrease in mRNA stability. This regulation is mediated by Ang II type 1 (AT1) receptors and abolished by NADPH oxidase inhibitors and dominant negative rac. The Ang II effect is also blocked by expression of superoxide dismutase (SOD), but not catalase, showing that superoxide is required. Dominant negative subunits, enzyme inhibitors and hydrogen peroxide experiments show that the apoptosis signal-regulating kinase 1 (ASK1)-p38 kinase pathway mediates downstream signaling from NADPH oxidase. Mechanical stretch also downregulates Kv4.3 3'-UTR reporter activity and this requires AT1 receptors and NADPH oxidase. Thus, activation of AT1 receptors by Ang II or stretch specifically destabilizes cardiac myocyte Kv4.3 channel mRNA by activating NADPH oxidase. These results link long-term control of cardiac K+ channel gene expression to a physiological reactive oxygen species signaling pathway.
Department of Pharmacology, University of Pittsburgh, Pittsburgh, PA 15261, USA.
Chaoming
C
Zhou
Chandra
C
Ziegler
Lori A
LA
Birder
Alexandre F R
AF
Stewart
Edwin S
ES
Levitan
D020413:3' Untranslated Regions / Q000187:drug effects / Q000235:genetics; D000804:Angiotensin II / Q000494:pharmacology*; D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004789:Enzyme Activation; D020224:Expressed Sequence Tags; D017930:Genes, Reporter; D006321:Heart / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D051379:Mice; D032342:Muscle Cells / Q000166:cytology / Q000502:physiology; D009206:Myocardium / Q000166:cytology; D019255:NADPH Oxidases / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051672:Shal Potassium Channels / Q000235:genetics*
D020413:3' Untranslated Regions; D012333:RNA, Messenger; D051672:Shal Potassium Channels; D000804:Angiotensin II; D019255:NADPH Oxidases