The steroid hormone 20-hydroxyecdysone induces phosphorylation and aggregation of stromal interacting molecule 1 for store-operated calcium entry
The steroid hormone 20-hydroxyecdysone induces phosphorylation and aggregation of stromal interacting molecule 1 for store-operated calcium entry
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类固醇激素 20-羟基蜕皮酮诱导基质相互作用分子 1 的磷酸化和聚集，以实现钙库操作的钙进入
DOI：
10.1074/jbc.ra119.008484
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发表时间：
2019-10-11
影响因子：
4.8
通讯作者：
Zhao, Xiao-Fan
中科院分区：
文献类型：
--
作者：
Chen, Cai-Hua;Di, Yu-Qin;Zhao, Xiao-Fan
基质相互作用分子1（STIM 1）的寡聚化促进钙库操纵的钙内流（SOCE）;然而，STIM 1聚集的机制尚不清楚。在这里，使用鳞翅目昆虫和农业害虫棉铃虫（Helicoverpa armigera）作为模型和免疫印迹，RT-qPCR，RNA干扰（RNAi）和ChIP测定，我们发现类固醇激素20-羟基蜕皮激素（20 E）通过G蛋白偶联受体（GPCR）和20 E核受体（EcRB 1）上调STIM 1的表达。我们还在STIM 1基因的5′上游区域鉴定了蜕皮激素反应元件（EcRE），并注意到STIM 1在变态期间位于幼虫中肠。STIM 1敲低延迟化蛹时间，防止中肠重塑，并减少20 E诱导的基因转录。STIM 1基因敲除棉铃虫表皮细胞系HaEpi抑制20 E诱导的钙离子内流和凋亡。此外，20 E诱导STIM 1聚集到斑点和易位向细胞膜。GPCR抑制剂、磷脂酶C（PLC）和三磷酸肌醇受体（IP 3 R）抑制20 E诱导的STIM 1磷酸化，我们发现两个GPCR参与20 E诱导的STIM 1磷酸化。20 E通过蛋白激酶C（PKC）诱导Ser-485上的STIM 1磷酸化，我们观察到Ser-485磷酸化对于STIM 1聚集、与钙释放激活的钙通道调节剂1（Orai 1）相互作用、钙离子内流和20 E诱导的细胞凋亡至关重要。这些结果表明，20 E上调STIM 1磷酸化，通过GPCR聚集，然后与Orai 1相互作用，诱导SOCE，从而促进昆虫变态期间中肠的凋亡。
Oligomerization of stromal interacting molecule 1 (STIM1) promotes store-operated calcium entry (SOCE); however, the mechanism of STIM1 aggregation is unclear. Here, using the lepidopteran insect and agricultural pest cotton bollworm (Helicoverpa armigera) as a model and immunoblotting, RT-qPCR, RNA interference (RNAi), and ChIP assays, we found that the steroid hormone 20-hydroxyecdysone (20E) up-regulates STIM1 expression via G protein–coupled receptors (GPCRs) and the 20E nuclear receptor (EcRB1). We also identified an ecdysone-response element (EcRE) in the 5′-upstream region of the STIM1 gene and also noted that STIM1 is located in the larval midgut during metamorphosis. STIM1 knockdown in larvae delayed pupation time, prevented midgut remodeling, and decreased 20E-induced gene transcription. STIM1 knockdown in a H. armigera epidermal cell line, HaEpi, repressed 20E-induced calcium ion influx and apoptosis. Moreover, 20E-induced STIM1 clustering to puncta and translocation toward the cell membrane. Inhibitors of GPCRs, phospholipase C (PLC), and inositol trisphosphate receptor (IP3R) repressed 20E-induced STIM1 phosphorylation, and we found that two GPCRs are involved in 20E-induced STIM1 phosphorylation. 20E-induced STIM1 phosphorylation on Ser-485 through protein kinase C (PKC), and we observed that Ser-485 phosphorylation is critical for STIM1 clustering, interaction with calcium release-activated calcium channel modulator 1 (Orai1), calcium ion influx, and 20E-induced apoptosis. These results suggest that 20E up-regulates STIM1 phosphorylation for aggregation via GPCRs, followed by interaction with Orai1 to induce SOCE, thereby promoting apoptosis in the midgut during insect metamorphosis.