Selective loss of fine tuning of Gq/11 signaling by RGS2 protein exacerbates cardiomyocyte hypertrophy

被引:88
作者
Zhang, W
Anger, T
Su, JL
Hao, JM
Xu, XM
Zhu, M
Gach, A
Cui, L
Liao, RL
Mende, U
机构
[1] Brigham & Womens Hosp, Div Cardiovasc, Boston, MA 02115 USA
[2] Harvard Univ, Sch Med, Boston, MA 02115 USA
关键词
D O I
10.1074/jbc.M507871200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Alterations in cardiac G protein- mediated signaling, most prominently G(q/ 11) signaling, are centrally involved in hypertrophy and heart failure development. Several RGS proteins that can act as negative regulators of G protein signaling are expressed in the heart, but their functional roles are still poorly understood. RGS expression changes have been described in hypertrophic and failing hearts. In this study, we report a marked decrease in RGS2 ( but not other major cardiac RGS proteins ( RGS3 - RGS5)) that occurs prior to hypertrophy development in different models with enhanced G(q/11) signaling ( transgenic expression of activated G alpha(q)* and pressure overload due to aortic constriction). To assess functional consequences of selective down- regulation of endogenous RGS2, we identified targeting sequences for effective RGS2 RNA interference and used lipid- based transfection to achieve uptake of fluorescently labeled RGS2 small interfering RNA in > 90% of neonatal and adult ventricular myocytes. Endogenous RGS2 expression was dose- dependently suppressed ( up to 90%) with no major change in RGS3 RGS5. RGS2 knockdown increased phenylephrine- and endothelin-1- induced phospholipase C beta stimulation in both cell types and exacerbated the hypertrophic effect ( increase in cell size and radiolabeled protein) in neonatal myocytes, with no major change in G(q/11)- mediated ERK1/ 2, p38, or JNK activation. Taken together, this study demonstrates that endogenous RGS2 exerts functionally important inhibitory restraint on G(q/ 11)- mediated phospholipase C beta activation and hypertrophy in ventricular myocytes. Our findings point toward a potential pathophysiological role of loss of fine tuning due to selective RGS2 down- regulation in G(q/ 11)- mediated remodeling. Furthermore, this study shows the feasibility of effective RNA interference in cardiomyocytes using lipid- based small interfering RNA transfection.
引用
收藏
页码:5811 / 5820
页数:10
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