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| Content Provider | Journal of Biological Chemistry (JBC) |
|---|---|
| Author | Ikari, Akira Tonegawa, Chie Sanada, Ayumi Kimura, Toru Sakai, Hideki Hayashi, Hisayoshi Hasegawa, Hajime Yamaguchi, Masahiko Yamazaki, Yasuhiro Endo, Satoshi Matsunaga, Toshiyuki Sugatani, Junko |
| Abstract | Claudin-16 (CLDN16) regulates the paracellular reabsorption of Mg2+ in the thick ascending limb of Henle's loop. However, the mechanism regulating the tight junctional localization of CLDN16 remains unknown. In yeast two-hybrid systems, we found that CLDN16 bound to syntaxin 8 (STX8), a target soluble N-ethylmaleimide-sensitive factor attachment protein receptor. We have examined the effect of STX8 on the localization and function of CLDN16 using Madin-Darby canine kidney cells expressing FLAG-tagged CLDN16. A pulldown assay showed that the carboxyl cytoplasmic region of human CLDN16 bound to STX8. CLDN16 was localized in the thick ascending limb, whereas STX8 was widely distributed throughout the rat kidney. An association between CLDN16 and STX8 was observed in rat renal homogenates and Madin-Darby canine kidney cells. STX8 siRNA decreased the cell surface localization of CLDN16 and transepithelial electrical resistance and permeability to Mg2+ but increased the co-localization of CLDN16 with early endosome and lysosome markers. Dephosphorylation of CLDN16 by protein kinase A inhibitors and S217A mutant, a dephosphorylated form, decreased the association with STX8 and the cell surface localization of CLDN16. Recycling assays indicated that STX8 siRNA decreased the trafficking of CLDN16 to the plasma membrane without affecting endocytosis. Dominant negative Rab11 and recycling inhibitor primaquine decreased the cell surface localization of CLDN16, which was similar to that in STX8 siRNA-transfected cells. These results suggest that STX8 mediates the recycling of CLDN16 and constitutes an important component of the CLDN16 trafficking machinery in the kidney. |
| Related Links | http://www.jbc.org/content/289/19/13112.abstract |
| Ending Page | 13123 |
| Starting Page | 13112 |
| Page Count | 12 |
| File Format | HTM / HTML PDF |
| ISSN | 00219258 |
| Journal | Journal of Biological Chemistry (JBC) |
| Issue Number | 19 |
| Volume Number | 289 |
| DOI | 10.1074/jbc.M113.541193 |
| e-ISSN | 1083351X |
| Language | English |
| Publisher | American Society for Biochemistry and Molecular Biology |
| Publisher Date | 2014-05-09 |
| Access Restriction | Open |
| Subject Keyword | Confocal Microscopy Kidney Magnesium Homeostasis Tight Junctions Trafficking Claudin Syntaxin Membrane Biology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Biochemistry Molecular Biology |
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