The C-Terminal Region of Serotonin Transporter Is Important for Its Trafficking and Glycosylation
We investigated the effects of brefeldin A and ilimaquinone, inhibitors of membrane trafficking, using serotonin transporter (SERT)–expressing COS-7 cells. Both drugs significantly inhibited the serotonin uptake activity of SERT and caused SERT to be retained in the endoplasmic reticulum (ER), indic...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2009-01-01
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| Series: | Journal of Pharmacological Sciences |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1347861319310758 |
| _version_ | 1819059458286813184 |
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| author | Mariko Nobukuni Hideki Mochizuki Satoshi Okada Nobuhisa Kameyama Anna Tanaka Hikaru Yamamoto Taku Amano Takahiro Seki Norio Sakai |
| author_facet | Mariko Nobukuni Hideki Mochizuki Satoshi Okada Nobuhisa Kameyama Anna Tanaka Hikaru Yamamoto Taku Amano Takahiro Seki Norio Sakai |
| author_sort | Mariko Nobukuni |
| collection | DOAJ |
| description | We investigated the effects of brefeldin A and ilimaquinone, inhibitors of membrane trafficking, using serotonin transporter (SERT)–expressing COS-7 cells. Both drugs significantly inhibited the serotonin uptake activity of SERT and caused SERT to be retained in the endoplasmic reticulum (ER), indicating that membrane trafficking is an important factor for SERT functional regulation. In agreement with previous reports, a C-terminal–deletion mutant of SERT (SERTΔCT) mostly localized to the ER and completely lacked serotonin uptake activity. To further elucidate the role of the C-terminus of SERT, we investigated whether overexpression of FLAG-tagged SERT C-terminus (FLAG-SERT-CT) affected the serotonin uptake activity and glycosylation of SERT. Interestingly, when concomitantly expressed with full-length FLAG-SERT in COS-7 cells, FLAG-SERT-CT increased the serotonin uptake activity and mature glycosylation of FLAG-SERT. These results indicate that the C-terminal region of SERT plays a crucial role in the functional regulation of SERT via membrane trafficking and glycosylation. In addition, proteasome inhibitors induced apparent ER stress, significantly decreased the serotonin uptake activity and mature glycosylation of SERT and caused SERT to be localized to the ER, suggesting that SERT function would be attenuated via membrane trafficking in pathological states that trigger ER stress. Keywords:: serotonin transporter, membrane trafficking, glycosylation, endoplasmic reticulum stress, plasma membrane |
| first_indexed | 2024-12-21T14:11:25Z |
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| institution | Directory Open Access Journal |
| issn | 1347-8613 |
| language | English |
| last_indexed | 2024-12-21T14:11:25Z |
| publishDate | 2009-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Pharmacological Sciences |
| spelling | doaj.art-8b3edcb3627f4851a9fcd9e4c3ec69782022-12-21T19:01:01ZengElsevierJournal of Pharmacological Sciences1347-86132009-01-011114392404The C-Terminal Region of Serotonin Transporter Is Important for Its Trafficking and GlycosylationMariko Nobukuni0Hideki Mochizuki1Satoshi Okada2Nobuhisa Kameyama3Anna Tanaka4Hikaru Yamamoto5Taku Amano6Takahiro Seki7Norio Sakai8Departments of Molecular and Pharmacological Neuroscience and 1-2-3 Kasumi Minami-ku, Hiroshima 734-8551, JapanOphthalmology and Visual Sciences, Graduate School of Biomedical Sciences, Hiroshima University, 1-2-3 Kasumi Minami-ku, Hiroshima 734-8551, JapanDepartments of Molecular and Pharmacological Neuroscience and 1-2-3 Kasumi Minami-ku, Hiroshima 734-8551, JapanDepartments of Molecular and Pharmacological Neuroscience and 1-2-3 Kasumi Minami-ku, Hiroshima 734-8551, JapanDepartments of Molecular and Pharmacological Neuroscience and 1-2-3 Kasumi Minami-ku, Hiroshima 734-8551, JapanDepartments of Molecular and Pharmacological Neuroscience and 1-2-3 Kasumi Minami-ku, Hiroshima 734-8551, JapanDepartments of Molecular and Pharmacological Neuroscience and 1-2-3 Kasumi Minami-ku, Hiroshima 734-8551, JapanDepartments of Molecular and Pharmacological Neuroscience and 1-2-3 Kasumi Minami-ku, Hiroshima 734-8551, JapanDepartments of Molecular and Pharmacological Neuroscience and 1-2-3 Kasumi Minami-ku, Hiroshima 734-8551, Japan; Corresponding author. nsakai@hiroshima-u.ac.jpWe investigated the effects of brefeldin A and ilimaquinone, inhibitors of membrane trafficking, using serotonin transporter (SERT)–expressing COS-7 cells. Both drugs significantly inhibited the serotonin uptake activity of SERT and caused SERT to be retained in the endoplasmic reticulum (ER), indicating that membrane trafficking is an important factor for SERT functional regulation. In agreement with previous reports, a C-terminal–deletion mutant of SERT (SERTΔCT) mostly localized to the ER and completely lacked serotonin uptake activity. To further elucidate the role of the C-terminus of SERT, we investigated whether overexpression of FLAG-tagged SERT C-terminus (FLAG-SERT-CT) affected the serotonin uptake activity and glycosylation of SERT. Interestingly, when concomitantly expressed with full-length FLAG-SERT in COS-7 cells, FLAG-SERT-CT increased the serotonin uptake activity and mature glycosylation of FLAG-SERT. These results indicate that the C-terminal region of SERT plays a crucial role in the functional regulation of SERT via membrane trafficking and glycosylation. In addition, proteasome inhibitors induced apparent ER stress, significantly decreased the serotonin uptake activity and mature glycosylation of SERT and caused SERT to be localized to the ER, suggesting that SERT function would be attenuated via membrane trafficking in pathological states that trigger ER stress. Keywords:: serotonin transporter, membrane trafficking, glycosylation, endoplasmic reticulum stress, plasma membranehttp://www.sciencedirect.com/science/article/pii/S1347861319310758 |
| spellingShingle | Mariko Nobukuni Hideki Mochizuki Satoshi Okada Nobuhisa Kameyama Anna Tanaka Hikaru Yamamoto Taku Amano Takahiro Seki Norio Sakai The C-Terminal Region of Serotonin Transporter Is Important for Its Trafficking and Glycosylation Journal of Pharmacological Sciences |
| title | The C-Terminal Region of Serotonin Transporter Is Important for Its Trafficking and Glycosylation |
| title_full | The C-Terminal Region of Serotonin Transporter Is Important for Its Trafficking and Glycosylation |
| title_fullStr | The C-Terminal Region of Serotonin Transporter Is Important for Its Trafficking and Glycosylation |
| title_full_unstemmed | The C-Terminal Region of Serotonin Transporter Is Important for Its Trafficking and Glycosylation |
| title_short | The C-Terminal Region of Serotonin Transporter Is Important for Its Trafficking and Glycosylation |
| title_sort | c terminal region of serotonin transporter is important for its trafficking and glycosylation |
| url | http://www.sciencedirect.com/science/article/pii/S1347861319310758 |
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