Abstract
The effects of angiotensin II (Ang II) are mediated primarily by Ang II type 1 receptors, which in turn are coupled to heterotrimeric G proteins. After receptor activation, the Gα and Gβγ subunits dissociate, contributing to the signaling cascades involving protein kinase C (PKC) activation. Regulators of G protein signaling (RGS proteins) comprise a class of proteins that have been shown to negatively regulate the Gα subunit. We examined which RGS sequences were expressed in vascular smooth muscle cells and which of these were regulated by Ang II. Reverse transcription-polymerase chain reaction showed that of 16 RGS sequences screened, six RGS transcripts (RGS2, 3, 10, 11, and 12 and GAIP) were present. Northern blot analysis demonstrated that RGS3, 10, and 12 and GAIP were not regulated by Ang II at the mRNA level. In contrast, RGS2 mRNA was rapidly and dose dependently increased (395 ± 24% peak, 45 min) by Ang II but returned to baseline level by 6 to 8 h. Phorbol-12-myristate-13-acetate, a PKC activator, robustly increased RGS2. This signal was attenuated by the PKC inhibitor GF 109203X (50 ± 4%) and by phorbol-12,13-dibutyrate-mediated down-regulation of PKC (48 ± 13%). Tyrosine kinase inhibition and calcium deprivation did not affect the up-regulation of RGS2 mRNA after Ang II stimulation. Actinomycin D treatment inhibited both Ang II- and phorbol-12-myristate-13-acetate-stimulated RGS2 up-regulation, suggesting activation of transcription by these agonists. The stability of RGS2 mRNA did not appear to be affected by Ang II. Thus, RGS2 is a likely candidate for negative regulation of the G proteins coupled to the Ang II type 1 receptor in vascular smooth muscle cells. Regulation of this protein may be of critical importance in modulating the role of Ang II in vascular disease.
Footnotes
- Received May 7, 1999.
- Accepted December 3, 1999.
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Send reprint requests to: Sharon Grant, Ph.D., Division of Cardiology, Emory University, Suite 319 WMB, 1639 Pierce Dr., Atlanta, GA 30322. E-mail: slgrant{at}emory.edu
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↵1 These authors contributed equally to this work.
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This work was supported by National Institutes of Health Grant HL60728. It was presented in abstract form at Experimental Biology 99, Washington, DC, April 1999 [Grant SL, Griendling KK, Lassègue B, Ushio-Fukai M and Alexander RW (1999) Angiotensin II regulates RGS expression in vascular smooth muscle cells].
- The American Society for Pharmacology and Experimental Therapeutics
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