Cx43 phosphorylation on S279/282 and intercellular communication are regulated by IP3/IP3 receptor signaling.
Cx43 phosphorylation on S279/282 and intercellular communication are regulated by IP3/IP3 receptor signaling.
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S279/282 上的 Cx43 磷酸化和细胞间通讯受 IP3/IP3 受体信号传导调节。
DOI：
10.1186/s12964-014-0058-6
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发表时间：
2014-09-28
期刊：
影响因子：
--
通讯作者：
Luo D
中科院分区：
文献类型：
--
作者：
Kang M;Lin N;Li C;Meng Q;Zheng Y;Yan X;Deng J;Ou Y;Zhang C;He J;Luo D
肌醇1，4，5-三磷酸受体（IP 3R）在多种细胞类型的Ca 2+释放过程中起关键作用。此外，IP 3R分布在心室闰盘中，但其在该特定部位的功能尚不清楚。缝隙连接蛋白（Cx43）是心室肌中主要的间隙连接蛋白，通过多个残基的磷酸化（去磷酸化）参与多种信号通路的调节。在这里，我们研究了IP 3R在细胞间通讯中的调节作用及其机制。在新生大鼠和成年小鼠心室肌细胞IP 3R共定位和免疫共沉淀与Cx43在GJ斑块检测免疫染色和蛋白质印迹分析。用拮抗剂阻断IP 3R或用shRNA沉默pan-IP 3R表达阻碍了6-羧基荧光素（6-CFDA）通过GJ的扩散，并在培养的融合新生心肌细胞中抑制了Ca 2+瞬变，而用IP 3酯或ATP刺激IP 3R产生了相反的效果。同样地，通过GJ的6-CFDA传播通过分离的成体心肌细胞的细胞对中的IP 3R活化或抑制来调节。此外，IP 3R激活或IP 3R抑制分别促进或抑制S279/282上的Cx43磷酸化。定点突变表明，突变体Cx43-S282 A（丙氨酸）的表达抑制S279/282磷酸化和GJ通透性，而S279 A突变体在心室肌细胞中表现出相反的效果。这些突变体在HEK 293细胞中的表达显示，具有S279/282双突变的细胞不能表达外源性Cx43，而具有S279或S282单突变的细胞显示Cx43过表达，增加S279/282的磷酸化和促进细胞间通讯。这些结果首次表明，IP 3R与Cx43物理相互作用，并参与调节S279/282上的Cx43磷酸化，从而影响心室肌细胞中的GJ细胞间通讯。本文的在线版本（doi：10.1186/s12964-014-0058-6）包含补充材料，可供授权用户使用。
Inositol 1,4,5-trisphosphate receptor (IP3R) plays a pivotal role in the Ca2+ release process in a variety of cell types. Additionally, IP3R is distributed in ventricular intercalated discs, but its function(s) in this particular site remains unknown. Connexin (Cx43), the predominant gap junction (GJ) protein in ventricular myocardium, is linked to several signaling pathways that regulate Cx43 properties by (de)phosphorylation on multiple residues. Here, we investigated the regulatory role of IP3R in cell-cell communication and the mechanism(s) underlying this effect. In neonatal rat and adult mouse ventricular myocytes IP3R co-localized and co-immunoprecipitated with Cx43 in GJ plaques detected by immunostaining and western blot assays. Blocking IP3R with antagonists or silencing pan-IP3R expression with shRNA hindered the 6-carboxyfluorescein (6-CFDA) diffusion through GJs and desynchronized Ca2+ transients among confluent neonatal myocytes in culture, whereas stimulation of IP3R with IP3 ester or ATP exerted the opposite effect. Likewise, 6-CFDA propagation through GJs was modulated by IP3R activation or inhibition in cell pairs of isolated adult cardiomyocytes. Furthermore, IP3R activation or IP3R suppression promoted or suppressed, respectively, Cx43 phosphorylation on S279/282. Site-directed mutagenesis indicated that expression of a mutant Cx43-S282A (alanine) inhibited S279/282 phosphorylation and GJ permeability, while the S279A mutant showed the opposite effect in ventricular myocytes. Expression of these mutants in HEK293 cells revealed that cells with a dual S279/282 mutation failed to express exogenous Cx43, whereas cells with a single S279 or S282 mutation displayed Cx43 overexpression with increased phosphorylation of S279/282 and promotion of intercellular communication. These results demonstrated, for the first time, that IP3R physically interacts with Cx43 and participates in the regulation of Cx43 phosphorylation on S279/282, thereby affecting GJ intercellular communication in ventricular myocytes. The online version of this article (doi:10.1186/s12964-014-0058-6) contains supplementary material, which is available to authorized users.
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影响因子：
--
作者：
Evans, W. Howard;Leybaert, Luc
通讯作者：
Leybaert, Luc
影响因子：
10.8
作者：
Bruce AF;Rothery S;Dupont E;Severs NJ
通讯作者：
Severs NJ
影响因子：
20.1
作者：
Kizana, Eddy;Chang, Connie Y.;Marban, Eduardo
通讯作者：
Marban, Eduardo
DOI：
10.3109/15419060009109019
发表时间：
2000-01-01
期刊：
CELL ADHESION AND COMMUNICATION
影响因子：
--
作者：
Opsahl, H;Rivedal, E
通讯作者：
Rivedal, E
影响因子：
5.5
作者：
Jansen, John A.;Noorman, Maartje;Musa, Hassan;Stein, Mera;de Jong, Sanne;van der Nagel, Roel;Hund, Thomas J.;Mohler, Peter J.;Vos, Marc A.;van Veen, Toon A.;de Bakker, Jacques M.;Delmar, Mario;van Rijen, Harold V.
通讯作者：
van Rijen, Harold V.