Calsequestrin-1 Regulates Store-Operated Ca2+ Entry by Inhibiting STIM1 Aggregation
Calsequestrin-1 Regulates Store-Operated Ca2+ Entry by Inhibiting STIM1 Aggregation
复制标题
Calsequestrin-1 通过抑制 STIM1 聚集来调节存储操作的 Ca2 进入
DOI：
10.1159/000445574
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发表时间：
2016-05
影响因子：
--
通讯作者：
Luo Dali
中科院分区：
文献类型：
--
作者：
Zhang Lane;Wang Limin;Li Shu;Xue Jingyi;Luo Dali
背景/目的：基质相互作用分子-1 （STIM1）聚集并重新分布到质膜上与Orai1相互作用是储存操作Ca2+进入（SOCE）的核心机制。先前的研究发现calsequestrin-1 （CSQ1）通过与STIM1相互作用并抑制STIM1/Orai1相互作用调控HEK293细胞的SOCE。在这里，我们进一步研究CSQ1/STIM1相互作用如何影响SOCE。方法：在转染不同CSQ1突变体的HeLa细胞中，使用共聚焦显微镜，用Imaris软件测量和量化经thapsigargin耗尽Ca2+存储后STIM1与CSQ1或Orai1的聚集和共定位。通过共免疫沉淀和Fura2分别检测CSQ1/STIM1和STIM1/Orai1的相互作用以及内部Ca2+的变化。结果：Wt-CSQ1过表达显著减少了STIM1在膜周和细胞质区域的聚集，而c端氨基酸362-396缺失突变体（C35）的过表达则没有。同样，在wt-CSQ1中观察到SOCE的显著降低，CSQ1单体化和CSQ1/STIM1相互作用的增加，以及STIM1/Orai1关联的降低，而在c35转染的细胞中则没有观察到。此外，缺乏c端AA 388-396缺失的突变体在抑制STIM1聚集和与Orai1关联方面的作用弱于wt-CSQ1。结论：我们的研究结果表明，CSQ1单体通过与STIM1相互作用并通过其c端氨基酸362-396减弱STIM1聚集来抑制SOCE。
Background/Aims: Stromal interacting molecule-1 (STIM1) aggregation and redistribution to plasma membrane to interact with Orai1 constitute the core mechanism of store-operated Ca2+ entry (SOCE). Previous study has revealed that calsequestrin-1 (CSQ1) regulates SOCE in HEK293 cells through interacting with STIM1 and inhibiting STIM1/Orai1 interaction. Here, we further investigate how CSQ1/STIM1 interaction affects SOCE. Methods: Using confocal microscopy, STIM1 aggregation and co-localizations with CSQ1 or Orai1 upon Ca2+ store depletion by thapsigargin were measured and quantified by Imaris software in HeLa cells transfected with different CSQ1 mutants. The interactions of CSQ1/STIM1 and STIM1/Orai1, and internal Ca2+ changes were detected by co-immunoprecipitation and Fura2, respectively. Results: Wt-CSQ1 overexpression significantly reduced STIM1 clustering in the perimembrane and cytosolic regions, whereas over-expression of a C-terminal amino acid 362-396 deletion mutant (C35) did not. Consistently, a significant depression of SOCE, increased CSQ1 monomerization and CSQ1/STIM1 interaction, and a reduced STIM1/Orai1 association were observed in wt-CSQ1 but not in C35-transfected cells. Additionally, mutant lacking C-terminal AA 388-396 deletion exerted weaker potency in inhibiting STIM1 aggregation and association with Orai1 than wt-CSQ1. Conclusions: Our results demonstrate that CSQ1 monomers suppress SOCE by interacting with STIM1 and attenuating STIM1 aggregation via its C-terminal amino acid 362-396.
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影响因子：
64.5
作者：
Park CY;Hoover PJ;Mullins FM;Bachhawat P;Covington ED;Raunser S;Walz T;Garcia KC;Dolmetsch RE;Lewis RS
通讯作者：
Lewis RS
影响因子：
5.3
作者：
Smyth JT;Hwang SY;Tomita T;DeHaven WI;Mercer JC;Putney JW
通讯作者：
Putney JW
影响因子：
3.4
作者：
Györke, I;Hester, N;Györke, S
通讯作者：
Györke, S
影响因子：
64.8
作者：
Prakriya, Murali;Feske, Stefan;Hogan, Patrick G.
通讯作者：
Hogan, Patrick G.
影响因子：
3.4
作者：
R. Weerasekera;Y. She;Kelly A. Markham;Yu Bai;N. Opalka;S. Orlicky;F. Sicheri;T. Kislinger;G. Schmitt-Ulms
通讯作者：
R. Weerasekera;Y. She;Kelly A. Markham;Yu Bai;N. Opalka;S. Orlicky;F. Sicheri;T. Kislinger;G. Schmitt-Ulms