Modulation of angiotensin II receptor (AT2) mRNA levels in R3T3 cells.
R3T3 cells, a mouse fibroblast cell line, express the type 2 angiotensin II receptor (AT2), but not the AT1 subtype. We previously reported that expression of AT2 sites in these cells were regulated by various conditions: 1. The number of AT2 sites increased considerably when cells were contact-inhibited; 2. Stimulation of R3T3 cells with various mitogens caused a rapid decline of AT2 binding sites; and 3. Stimulation of cells with angiotensin ligands resulted in upregulation of the AT2 sites. In this study, to determine if altered AT2 expression is under transcriptional, posttranscriptional, or translational control, we examined the level of AT2 mRNA in R3T3 cells in response to various treatments. There was a 200-fold increase in AT2 mRNA levels in quiescent cells as compared to growing cells. Results from nuclear run-on assays suggested that the differences in AT2 mRNA levels were primarily caused by changes in the rate of AT2 gene transcription. Stimulation of cells with fibroblast growth factor caused an approximate threefold reduction of AT2 mRNA levels, and also increased the rate of degradation of AT2 mRNA, which correlated with the decrease in AT2 binding activity seen under these conditions. However, whereas treatment with angiotensin ligands increased AT2 binding activity, the level of AT2 transcripts did not increase. This pattern of expression implies that regulation of AT2 receptors occurs at multiple levels, involving translational and/or posttranslational as well as transcriptional control, and further affords the cell the ability to rapidly modulate the number of AT2 binding sites in response to changing extracellular conditions.
Camp
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Department of Signal Transduction, Parke-Davis Pharmaceutical Research Division, Warner-Lambert Co., Ann Arbor, MI 48105, USA.
Dudley
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D000804:Angiotensin II / Q000378:metabolism*; D000818:Animals; D015152:Blotting, Northern; D002448:Cell Adhesion; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D003001:Cloning, Molecular; D005346:Fibroblast Growth Factors / Q000494:pharmacology; D005347:Fibroblasts; D008024:Ligands; D051379:Mice; D008934:Mitogens / Q000494:pharmacology; D011485:Protein Binding; D014176:Protein Biosynthesis; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000378:metabolism*; D011945:Receptors, Angiotensin / Q000096:biosynthesis* / Q000235:genetics; D014158:Transcription, Genetic; D015854:Up-Regulation / Q000187:drug effects
D008024:Ligands; D008934:Mitogens; D012333:RNA, Messenger; D011945:Receptors, Angiotensin; D000804:Angiotensin II; D005346:Fibroblast Growth Factors