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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 李芳仁 | |
| dc.contributor.author | Pei-Chin Tsai | en |
| dc.contributor.author | 蔡佩秦 | zh_TW |
| dc.date.accessioned | 2021-06-13T01:14:26Z | - |
| dc.date.available | 2016-10-05 | |
| dc.date.copyright | 2011-10-05 | |
| dc.date.issued | 2011 | |
| dc.date.submitted | 2011-08-03 | |
| dc.identifier.citation | Alder-Baerens N, Lisman Q, Luong L, Pomorski T, Holthuis JC (2006) Loss of P4 ATPases Drs2p and Dnf3p disrupts aminophospholipid transport and asymmetry in yeast post-Golgi secretory vesicles. Mol Biol Cell 17: 1632-1642
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/29674 | - |
| dc.description.abstract | Drs2p 為一類的P 型三磷酸腺脢，位在高機氏體上且具有翻轉磷脂質絲胺酸(PS)
的能力。Gea2p 為腺嘌呤核苷二磷酸核醣化因子交換因子，會與Drs2p 結合並且 影響Drs2p 翻轉酶的活性。本文我們發現第一腺嘌呤核苷二磷酸核醣化因子相 似蛋白(Arl1p)會和Gea2p 共同去調控Drs2p 的翻轉酶活性。我們發現Drs2p 和 Gea2p 都會去影響Arl1p 的下游基因Imh1p 在細胞中的分佈。Drs2p 會和鳥糞呤 核苷三磷酸 (GTP) 形式的Arl1p 結合，當Drs2p 無法與Arl1p 結合，會影響其翻 轉酶的活性。Gea2p 用利用其N 端與Arl1p 結合，當其無法與Arl1p 結合時，也 會影響高基氏體上磷酸絲氨酸的轉位，但是不會影響到Gea2p 腺嘌呤核苷二磷 酸核醣化因子交換因子的活性。此外，Drs2p 和Gea2p 的結合需要有Arl1p 的存 在。因此我們的研究發現在特定高基氏體位置的Drs2p 活性會被Arl1p 以及Gea2p 所調控並且影響到Imh1p 在高基氏體上的分佈。 | zh_TW |
| dc.description.abstract | Drs2p, a resident type 4 P-type ATPase (P4-ATPase), requires for a
phosphatidylserine (PS) flippase activity in the yeast trans Golgi network (TGN) and plays essential roles in protein transport in the secretory and endocytic pathways. The Arf activator Gea2p interacts with Drs2p and stimulates its flippase activity in yeast TGN. Here we show that the ARF-like (ARL) protein, Arl1p, acts with Gea2p to modulate Drs2p activity at the TGN. We found that gea2- and drs2-null mutants, like arl1-null, exhibits severe defects in recruitment of Imh1p to the Golgi. Arl1p directly interacts with N-terminus of Drs2p in a GTP-dependent manner. Deletion of the Arl1p-interacting region of Drs2p results in a significant decrease of its flippase activity. In addition, the active form of Arl1p directly interacts with N-terminus of Gea2p. Deletion of the Arl1p-interacting region GEA2 impaired PS translocation on the TGN membranes, but appears to keep its GEF function for Arf. Deletion of ARL1 impairs Gea2p-Drs2p interaction. Thus, we infer that subcellular spatial regulation of flippase Drs2p by Arl1p and Arf-GEF Gea2p controls membrane dynamics at the trans-Golgi network. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T01:14:26Z (GMT). No. of bitstreams: 1 ntu-100-D95448007-1.pdf: 7740938 bytes, checksum: 1394cf19c9467a9d1768f4d00228f32c (MD5) Previous issue date: 2011 | en |
| dc.description.tableofcontents | Abstract --------------------------------------------------4
中文摘要 ---------------------------------------------------5 Abbreviations ---------------------------------------------6 Introduction ----------------------------------------------7 Materials and Methods ------------------------------------25 Results I. Arl1p associates with the Gea2p and Drs2p -------------41 II. Drs2p and Gea2p may be the effector of Arl1p ---------44 III. Arl1p, Gea2p and Drs2p may form the complex ---------56 Discussion -----------------------------------------------58 Tables Table 1. Yeast strains used in this study ----------------69 Table 2. Primers used in this study ----------------------70 Figures Figure 1. The structural GDP/GTP cycle of ARF6 -----------71 Figure 2. Regulation of coated vesicle formation by Arf---72 Figure 3. A model for the Arl3p-Syt1p-Arl1p-Imh1p pathway-73 Figure 4. P4-ATPases in Saccharomyces cerevisiae----------74 Figure 5. Vesicle-mediated protein-transport pathways that require P-type ATPases of the Drs2p family----------------75 Figure 6. Gea2p interacts with active form of Arl1p-------76 Figure 7. Active form of Arl1p interacts with Gea2p and Drs2p --------------- 77 Figure 8. N-terminal region of Gea2p is required for the interaction withactive form of Arl1p----------------------78 Figure 9. Drs2p interacts with active form of Arl1p in vitro--------------------- 79 Figure 10. N-terminal region of Drs2p is essential for the interaction with Arl1p in vivo ---------------------------80 Figure 11. Drs2p or Gea2p partially co-localizes with Arl1p--------------------- 81 Figure 12. Drs2p is not required for proper localization of Arl1p---------------- 82 Figure 13. Arl1p did not affect the localization of GFP-Drs2--------------------- 83 Figure 14. Gea2p does not determine the localization of Arl1p------------------ 84 Figure 15. Drs2p and Gea2p are involved in the localization of Imh1p and Gas1p------------------------------------85 Figure 16. Miss-localization of Imh1p to the cytoplasm of DRS2 mutants---- 86 Figure 17. Arl1p but not Drs2p or Gea2p affects the subcellular distribution of GFP–Gga2.---------------------87 Figure 18. Interaction of Drs2p with Arl1p is required for Golgi localization of Imh1p-------------------------------88 Figure 19. N-terminal region of Gea2p is required for the localization of Imh1p in gea2 mutant cells----------------89 Figure 20. Gea2p and Arl1p interact functionally in yeast cells------------------ 90 Figure 21. Over-expression of Drs2dN170 partial suppress the cold sensitive growth defect of drs2 cells----------------91 Figure 22. NBD-PS flippase activity in TGN membranes requires Arl1p------ 92 Figure 23. Active form of Arl1p is required for stimulation of NBD-PS flippase activity in TGN membranes--------------94 Figure 24. Loss of plasma membrane PS asymmetry in arl1-null cells--------- 95 Figure 25. N-terminal region of Drs2p is required for the activity of Drs2p---- 96 Figure 26. N-terminal region of Gea2p (1-89 AAs) is important for the activity of translocalization of NBD-PS at TGN-----------------------97 Figure 27. Arl1p, Drs2p or Gea2p is required for the interaction between the other proteins--------------------98 Figure 28. Model for the network interactions between Gea2p, Arl1p and Drs2p and mediate the Drs2p fillipase activity-100 Reference -----------------------------------------------101 | |
| dc.language.iso | en | |
| dc.title | 第一腺嘌呤核苷二磷酸核醣化因子相似蛋白在
囊泡運輸中的功能性探討 | zh_TW |
| dc.title | Characterization of ADP-Ribosylation Factor-Like
Protein 1 in vesicle trafficking | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 99-2 | |
| dc.description.degree | 博士 | |
| dc.contributor.oralexamcommittee | 陳瑞華,王昭雯,張智芬,鄧述諄 | |
| dc.subject.keyword | 第一腺嘌呤核苷,二磷酸核醣化因子相似蛋白,囊泡運輸, | zh_TW |
| dc.subject.keyword | Arl1p,vesicle trafficking, | en |
| dc.relation.page | 138 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2011-08-03 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 分子醫學研究所 | zh_TW |
| 顯示於系所單位： | 分子醫學研究所 | |
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