Enhanced angiotensin receptor type 1 mRNA degradation and induction of polyribosomal mRNA binding proteins by angiotensin II in vascular smooth muscle cells.
Stimulation of cultured rat thoracic aorta vascular smooth muscle cells (VSMCs) with 100 nM angiotensin II (Ang II) reduces angiotensin receptor type 1 (AT1-R) gene expression. mRNA levels are reduced to approximately 30% of control levels 4 hr after the addition of Ang II to the culture medium. The loss of mRNA remains sustained for up to 24 hr after the addition of Ang II. The half-life of the AT1-R mRNA is approximately 2 hr in cells treated with a single dose of 100 nM Ang II. This represents a 3-fold reduction from its half-life of 6 hr in nonstimulated cells, as assessed by treatment with 5,6-dichlorobenzimidazole or actinomycin D to block transcription. Thus, the AT1-R mRNA is moderately unstable in VSMC and destabilized further by treatment with Ang II. Ang II-induced AT1-R mRNA destabilization is prevented by pretreatment with transcriptional inhibitors or the protein synthesis inhibitor cycloheximide, suggesting that Ang II-induced AT1-R mRNA destabilization requires the induction of an unknown factor or factors that are postulated to mediate this effect. AT1-R mRNA levels decrease more rapidly in vitro from a polyribosomal fraction isolated from VSMC exposed for 2 hr to 100 nM Ang II compared with that from vehicle-treated cells, suggesting that polyribosomal-associated AT1-R mRNA is at least one site of action for the mRNA destabilization effect of Ang II. Ang II stimulation induces a complex of polyribosomal proteins that bind specifically in the distal 350 bases of the AT1-R mRNA. Regulation of mRNA stability accounts in part for modulation of AT1-R gene expression by Ang II in VSMCs, and Ang II-induced AT1-R mRNA polyribosomal binding proteins are associated with this phenomenon.
Nickenig
G
G
Department of Pharmacology, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
Murphy
T J
TJ
D000803:Angiotensin I / Q000378:metabolism*; D000804:Angiotensin II / Q000494:pharmacology*; D000818:Animals; D001013:Aorta, Thoracic / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D001345:Autoradiography; D002478:Cells, Cultured; D003513:Cycloheximide / Q000494:pharmacology; D004355:Drug Stability; D009131:Muscle, Smooth, Vascular / Q000187:drug effects* / Q000378:metabolism*; D011132:Polyribosomes / Q000187:drug effects / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism*; D012335:RNA, Ribosomal / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000096:biosynthesis*; D051381:Rats; D011945:Receptors, Angiotensin / Q000096:biosynthesis / Q000378:metabolism*; D013268:Stimulation, Chemical; D013347:Subcellular Fractions / Q000378:metabolism
D012333:RNA, Messenger; D012335:RNA, Ribosomal; D016601:RNA-Binding Proteins; D011945:Receptors, Angiotensin; D000804:Angiotensin II; D000803:Angiotensin I; D003513:Cycloheximide