Dissertation/ Thesis
Mechanisms of retrograde transport from Golgi apparatus to endoplasmic reticulum ; Mécanismes de transport rétrograde entre l’appareil de Golgi et le reticulum endoplasmique
| العنوان: | Mechanisms of retrograde transport from Golgi apparatus to endoplasmic reticulum ; Mécanismes de transport rétrograde entre l’appareil de Golgi et le reticulum endoplasmique |
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| المؤلفون: | Lian, Yen-Ling |
| المساهمون: | Biologie Cellulaire et Cancer, Institut Curie Paris -Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Université Paris sciences et lettres, Gaëlle Boncompain |
| المصدر: | https://theses.hal.science/tel-04080838 ; Cellular Biology. Université Paris sciences et lettres, 2022. English. ⟨NNT : 2022UPSLS067⟩. |
| بيانات النشر: | HAL CCSD |
| سنة النشر: | 2022 |
| مصطلحات موضوعية: | Intracellular trafficking, Golgi apparatus, Secretory pathway, Trafic intracellulaire, Appareil de Golgi, Voies de sécrétion, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology |
| الوصف: | Mammalian cells are characterized by the co-existence of multiple pathways, including anterograde and retrograde transport. The Golgi apparatus has a central role in processing and sorting cargos in the bi-directional trafficking. The Retention Using Selective Hooks (RUSH) system allows to synchronize the transport of cargos from the ER to downstream compartments and to systematically analyze the secretory routes. Owing to the interaction of streptavidin (Str) and streptavidin-binding peptide (SBP), the reporter protein can be retained in the ER and then released by the addition of biotin. However, biotin has a high affinity to streptavidin, impairing reversibility of the RUSH assay. With the Artificial Ligands of Streptavidin (ALiS), Golgi-to-ER retrograde transport can be monitored using RUSH upon their washout.The reversible RUSH assay was firstly set up to study two mechanisms of Golgi-to-ER retrograde transport, including the KDEL-mediated retrieval pathway and the glycosylation enzyme recycling pathway. Core streptavidin fused with the ER-retention signal (Str-KDEL), or with the invariant chain (Ii-Str) were used as hooks. Golgi-resident enzymes, ManII* (Mannosidase II)-SBP-EGFP or ST* (Sialyltransferase)-SBP-GFP served as Golgi-targeted RUSH reporters. Our kinetic analysis showed that the Golgi-to-ER transport of ManII* and ST* are both slower through the glycosylation enzyme recycling pathway than the KDEL-meditated retrieval pathway. To characterize the role of putative regulatory factors in KDEL-mediated retrograde transport, we performed RNAi experiments targeting to COG3 and Rab6. Our data showed that knockdown of COG3 resulted in delayed retrograde transport of ManII* and ST*, and the depletion of Rab6 led to impaired trafficking of ST*, consistent with the previous studies. Our data indicated that there are two distinct pathways regulating the retrograde transport of Golgi cargos, including KDEL-mediated ER retrieval and ER recycling of Golgi glycosylation enzymes.Lastly, we have generated ... |
| نوع الوثيقة: | doctoral or postdoctoral thesis |
| اللغة: | English |
| Relation: | NNT: 2022UPSLS067; tel-04080838; https://theses.hal.science/tel-04080838; https://theses.hal.science/tel-04080838/document; https://theses.hal.science/tel-04080838/file/Curie_LIAN_2022_archivage.pdf |
| الاتاحة: | https://theses.hal.science/tel-04080838 https://theses.hal.science/tel-04080838/document https://theses.hal.science/tel-04080838/file/Curie_LIAN_2022_archivage.pdf |
| Rights: | info:eu-repo/semantics/OpenAccess |
| رقم الانضمام: | edsbas.7DB83B84 |
| قاعدة البيانات: | BASE |
| الوصف غير متاح. |