Type 3 Inositol 1,4,5-Trisphosphate Receptor is a Crucial Regulator of Calcium Dynamics Mediated by Endoplasmic Reticulum in HEK Cells
Being the largest the Ca<sup>2+</sup> store in mammalian cells, endoplasmic reticulum (ER)-mediated Ca<sup>2+</sup> signalling often involves both Ca<sup>2+</sup> release via inositol 1, 4, 5-trisphosphate receptors (IP<sub>3</sub>R) and store operated...
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MDPI AG
2020-01-01
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| Online Access: | https://www.mdpi.com/2073-4409/9/2/275 |
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| author | Lili Yue Liuqing Wang Yangchun Du Wei Zhang Kozo Hamada Yoshifumi Matsumoto Xi Jin Yandong Zhou Katsuhiko Mikoshiba Donald L. Gill Shengcheng Han Youjun Wang |
| author_facet | Lili Yue Liuqing Wang Yangchun Du Wei Zhang Kozo Hamada Yoshifumi Matsumoto Xi Jin Yandong Zhou Katsuhiko Mikoshiba Donald L. Gill Shengcheng Han Youjun Wang |
| author_sort | Lili Yue |
| collection | DOAJ |
| description | Being the largest the Ca<sup>2+</sup> store in mammalian cells, endoplasmic reticulum (ER)-mediated Ca<sup>2+</sup> signalling often involves both Ca<sup>2+</sup> release via inositol 1, 4, 5-trisphosphate receptors (IP<sub>3</sub>R) and store operated Ca<sup>2+</sup> entries (SOCE) through Ca<sup>2+</sup> release activated Ca<sup>2+</sup> (CRAC) channels on plasma membrane (PM). IP<sub>3</sub>Rs are functionally coupled with CRAC channels and other Ca<sup>2+</sup> handling proteins. However, it still remains less well defined as to whether IP<sub>3</sub>Rs could regulate ER-mediated Ca<sup>2+</sup> signals independent of their Ca<sup>2+</sup> releasing ability. To address this, we generated IP<sub>3</sub>Rs triple and double knockout human embryonic kidney (HEK) cell lines (IP<sub>3</sub>Rs-TKO, IP<sub>3</sub>Rs-DKO), and systemically examined ER Ca<sup>2+</sup> dynamics and CRAC channel activity in these cells. The results showed that the rate of ER Ca<sup>2+</sup> leakage and refilling, as well as SOCE were all significantly reduced in IP<sub>3</sub>Rs-TKO cells. And these TKO effects could be rescued by over-expression of IP<sub>3</sub>R3. Further, results showed that the diminished SOCE was caused by NEDD4L-mediated ubiquitination of Orai1 protein. Together, our findings indicate that IP<sub>3</sub>R3 is one crucial player in coordinating ER-mediated Ca<sup>2+</sup> signalling. |
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| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-12T06:42:01Z |
| publishDate | 2020-01-01 |
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| spelling | doaj.art-2a3aa1fc63af4c10b634f83cbab4a21f2023-09-03T00:57:12ZengMDPI AGCells2073-44092020-01-019227510.3390/cells9020275cells9020275Type 3 Inositol 1,4,5-Trisphosphate Receptor is a Crucial Regulator of Calcium Dynamics Mediated by Endoplasmic Reticulum in HEK CellsLili Yue0Liuqing Wang1Yangchun Du2Wei Zhang3Kozo Hamada4Yoshifumi Matsumoto5Xi Jin6Yandong Zhou7Katsuhiko Mikoshiba8Donald L. Gill9Shengcheng Han10Youjun Wang11Beijing Key Laboratory of Gene Resource and Molecular Development, College of Life Sciences, Beijing Normal University, Beijing 100875, ChinaBeijing Key Laboratory of Gene Resource and Molecular Development, College of Life Sciences, Beijing Normal University, Beijing 100875, ChinaBeijing Key Laboratory of Gene Resource and Molecular Development, College of Life Sciences, Beijing Normal University, Beijing 100875, ChinaDepartment of Molecular and Cellular Biology, College of Biological Science, University of Guelph, 50 Stone Rd E, Guelph, ON N1G 2W1, Canada Shanghai Institute for Advanced Immunochemical Studies, ShanghaiTech University, 393 Middle Huaxia Road, Shanghai 201210, ChinaCentral Institute for Experimental Animals, Kawasaki, Kanagawa 210-0821, JapanBeijing Key Laboratory of Gene Resource and Molecular Development, College of Life Sciences, Beijing Normal University, Beijing 100875, ChinaDepartment of Cellular and Molecular Physiology, Pennsylvania State University College of Medicine, Hershey, PA 17033, USA Shanghai Institute for Advanced Immunochemical Studies, ShanghaiTech University, 393 Middle Huaxia Road, Shanghai 201210, ChinaDepartment of Cellular and Molecular Physiology, Pennsylvania State University College of Medicine, Hershey, PA 17033, USABeijing Key Laboratory of Gene Resource and Molecular Development, College of Life Sciences, Beijing Normal University, Beijing 100875, ChinaBeijing Key Laboratory of Gene Resource and Molecular Development, College of Life Sciences, Beijing Normal University, Beijing 100875, ChinaBeing the largest the Ca<sup>2+</sup> store in mammalian cells, endoplasmic reticulum (ER)-mediated Ca<sup>2+</sup> signalling often involves both Ca<sup>2+</sup> release via inositol 1, 4, 5-trisphosphate receptors (IP<sub>3</sub>R) and store operated Ca<sup>2+</sup> entries (SOCE) through Ca<sup>2+</sup> release activated Ca<sup>2+</sup> (CRAC) channels on plasma membrane (PM). IP<sub>3</sub>Rs are functionally coupled with CRAC channels and other Ca<sup>2+</sup> handling proteins. However, it still remains less well defined as to whether IP<sub>3</sub>Rs could regulate ER-mediated Ca<sup>2+</sup> signals independent of their Ca<sup>2+</sup> releasing ability. To address this, we generated IP<sub>3</sub>Rs triple and double knockout human embryonic kidney (HEK) cell lines (IP<sub>3</sub>Rs-TKO, IP<sub>3</sub>Rs-DKO), and systemically examined ER Ca<sup>2+</sup> dynamics and CRAC channel activity in these cells. The results showed that the rate of ER Ca<sup>2+</sup> leakage and refilling, as well as SOCE were all significantly reduced in IP<sub>3</sub>Rs-TKO cells. And these TKO effects could be rescued by over-expression of IP<sub>3</sub>R3. Further, results showed that the diminished SOCE was caused by NEDD4L-mediated ubiquitination of Orai1 protein. Together, our findings indicate that IP<sub>3</sub>R3 is one crucial player in coordinating ER-mediated Ca<sup>2+</sup> signalling.https://www.mdpi.com/2073-4409/9/2/275ip<sub>3</sub>rstore-operated ca<sup>2+</sup> entryorai1sercanedd4lcalcium |
| spellingShingle | Lili Yue Liuqing Wang Yangchun Du Wei Zhang Kozo Hamada Yoshifumi Matsumoto Xi Jin Yandong Zhou Katsuhiko Mikoshiba Donald L. Gill Shengcheng Han Youjun Wang Type 3 Inositol 1,4,5-Trisphosphate Receptor is a Crucial Regulator of Calcium Dynamics Mediated by Endoplasmic Reticulum in HEK Cells Cells ip<sub>3</sub>r store-operated ca<sup>2+</sup> entry orai1 serca nedd4l calcium |
| title | Type 3 Inositol 1,4,5-Trisphosphate Receptor is a Crucial Regulator of Calcium Dynamics Mediated by Endoplasmic Reticulum in HEK Cells |
| title_full | Type 3 Inositol 1,4,5-Trisphosphate Receptor is a Crucial Regulator of Calcium Dynamics Mediated by Endoplasmic Reticulum in HEK Cells |
| title_fullStr | Type 3 Inositol 1,4,5-Trisphosphate Receptor is a Crucial Regulator of Calcium Dynamics Mediated by Endoplasmic Reticulum in HEK Cells |
| title_full_unstemmed | Type 3 Inositol 1,4,5-Trisphosphate Receptor is a Crucial Regulator of Calcium Dynamics Mediated by Endoplasmic Reticulum in HEK Cells |
| title_short | Type 3 Inositol 1,4,5-Trisphosphate Receptor is a Crucial Regulator of Calcium Dynamics Mediated by Endoplasmic Reticulum in HEK Cells |
| title_sort | type 3 inositol 1 4 5 trisphosphate receptor is a crucial regulator of calcium dynamics mediated by endoplasmic reticulum in hek cells |
| topic | ip<sub>3</sub>r store-operated ca<sup>2+</sup> entry orai1 serca nedd4l calcium |
| url | https://www.mdpi.com/2073-4409/9/2/275 |
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