Department of Pathology, University of Virginia Health System, Charlottesville, VA 22908, USA.
Hypertension. 2013 May;61(5):1021-7. doi: 10.1161/HYPERTENSIONAHA.111.00321. Epub 2013 Mar 18.
The G protein-coupled receptor kinase 4 (GRK4) negatively regulates the dopaminergic system by desensitizing the dopamine-1-receptor. The expressional control of GRK4 has not been reported, but here we show that the transcription factor c-Myc binds to the promoter of GRK4 and positively regulates GRK4 protein expression in human renal proximal tubule cells (RPTCs). Addition of phorbol esters to RPTCs not only increased c-Myc binding to the GRK4 promoter but also increased both phospho-c-Myc and GRK4 expression. The phorbol ester-mediated increase in GRK4 expression was completely blocked by the c-Myc inhibitor, 10074-G5, indicating that GRK4 is downstream of phospho-c-Myc. The autocrine production of angiotensin II (Ang II) in RPTCs increased the phosphorylation and activation of c-Myc and subsequently GRK4 expression. 3-Amino-4-thio-butyl sulfonate, an inhibitor of aminopeptidase A, increased RPTC secretion of Ang II. 3-Amino-4-thio-butyl sulfonate or Ang II increased the expression of both phospho-c-Myc and GRK4, which was blocked by 10074-G5. Blockade of the Ang II type 1 receptor with losartan decreased phospho-c-Myc and GRK4 expression. Both inhibition of c-Myc activity and blockade of Ang II type 1 receptor restored the coupling of dopamine-1-receptor to adenylyl cyclase stimulation in uncoupled RPTCs, whereas phorbol esters or Ang II caused the uncoupling of normally coupled RPTCs. We suggest that the Ang II type 1 receptor impairs dopamine-1-receptor function via c-Myc activation of GRK4. This novel pathway may be involved in the increase in blood pressure in hypertension that is mediated by increased activity of the renin-angiotensin system and decreased activity of the renal dopaminergic system.
G 蛋白偶联受体激酶 4(GRK4)通过使多巴胺-1-受体脱敏来负调控多巴胺能系统。尚未报道 GRK4 的表达调控,但我们在此表明转录因子 c-Myc 与 GRK4 启动子结合,并正向调节人肾近端小管细胞(RPTC)中的 GRK4 蛋白表达。佛波酯(Phorbol esters)的添加不仅增加了 c-Myc 与 GRK4 启动子的结合,而且还增加了磷酸化 c-Myc 和 GRK4 的表达。c-Myc 抑制剂 10074-G5 完全阻断了佛波酯介导的 GRK4 表达增加,表明 GRK4 是磷酸化 c-Myc 的下游产物。RPTC 中血管紧张素 II(Ang II)的自分泌产生增加了 c-Myc 的磷酸化和激活,随后增加了 GRK4 的表达。氨基肽酶 A 的抑制剂 3-氨基-4-硫代丁酸增加了 RPTC 分泌的 Ang II。3-氨基-4-硫代丁酸或 Ang II 增加了磷酸化 c-Myc 和 GRK4 的表达,这一表达被 10074-G5 阻断。用氯沙坦阻断 Ang II 型 1 受体降低了磷酸化 c-Myc 和 GRK4 的表达。抑制 c-Myc 活性和阻断 Ang II 型 1 受体都恢复了多巴胺-1-受体在未偶联的 RPTC 中与腺苷酸环化酶刺激的偶联,而佛波酯或 Ang II 则导致正常偶联的 RPTC 解偶联。我们认为 Ang II 型 1 受体通过 c-Myc 激活 GRK4 损害了多巴胺-1-受体功能。该新途径可能与高血压中血压升高有关,该升高是由肾素-血管紧张素系统活性增加和肾多巴胺能系统活性降低介导的。