Kidney Genetics Group, Academic Nephrology Unit, Henry Wellcome Laboratories for Medical Research,University of Sheffield Medical School, Sheffield, UK.
Hum Mol Genet. 2013 May 15;22(10):1924-39. doi: 10.1093/hmg/ddt031. Epub 2013 Feb 5.
Mutations in PKD1 (85%) or PKD2 (15%) account for almost all cases of autosomal dominant polycystic kidney disease (ADPKD). The ADPKD proteins, termed as polycystin-1 (PC1) and polycystin-2 (PC2), interact via their C-termini to form a receptor-ion channel complex whose function and regulation are not fully understood. Here, we report the first phosphorylated residue (Ser(829)) in PC2, whose dephosphorylation is mediated by PC1 binding through the recruitment of protein phosphatase-1 alpha (PP1α). Using a new phosphospecific antibody (pPC2) to this site, we demonstrate that Ser(829) is phosphorylated by Protein kinase A (PKA) but remains constitutively phosphorylated in cells and tissues lacking PC1. cAMP increased pSer(829) basolateral localization in MDCK cells in a time dependent manner and was essential for pronephric development in Xenopus embryos. When constitutively expressed, a complex phenotype associated with enhanced ATP-dependent ER Ca(2+) release and loss of growth suppression was observed in cycling cells. These results reveal a reciprocal functional link between PC1 and PC2 which is critically dependent on their interaction. Unopposed cAMP stimulated hyperphosphorylation of PC2 in the absence of functional PC1 could contribute to cyst initiation in PKD1 patients and represents a new molecular paradigm in understanding ADPKD pathogenesis.
PKD1(85%)或 PKD2(15%)的突变几乎占常染色体显性多囊肾病(ADPKD)的所有病例。ADPKD 蛋白,称为多囊蛋白-1(PC1)和多囊蛋白-2(PC2),通过它们的 C 末端相互作用形成一个受体离子通道复合物,其功能和调节尚不完全清楚。在这里,我们报告了 PC2 中的第一个磷酸化残基(Ser(829)),其去磷酸化是通过 PC1 结合通过募集蛋白磷酸酶-1α(PP1α)介导的。使用针对该位点的新磷酸化特异性抗体(pPC2),我们证明 Ser(829)被蛋白激酶 A(PKA)磷酸化,但在缺乏 PC1 的细胞和组织中仍然保持组成性磷酸化。cAMP 以时间依赖性方式增加 MDCK 细胞中 pSer(829)基底外侧定位,并且对非洲爪蟾胚胎的前肾发育是必需的。当组成性表达时,在有丝分裂细胞中观察到与增强的 ATP 依赖性内质网 Ca(2+)释放和生长抑制丧失相关的复杂表型。这些结果揭示了 PC1 和 PC2 之间的一种互惠功能联系,这严重依赖于它们的相互作用。在没有功能性 PC1 的情况下,cAMP 刺激的 PC2 过度磷酸化可能导致 PKD1 患者的囊肿起始,并代表了理解 ADPKD 发病机制的一个新的分子范例。