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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 劉雅雯(Ya-Wen Liu) | |
| dc.contributor.author | Julie Loh | en |
| dc.contributor.author | 駱怡君 | zh_TW |
| dc.date.accessioned | 2021-06-17T03:19:37Z | - |
| dc.date.available | 2018-08-01 | |
| dc.date.copyright | 2018-08-01 | |
| dc.date.issued | 2018 | |
| dc.date.submitted | 2018-06-26 | |
| dc.identifier.citation | Antonny, B., Burd, C., De Camilli, P., Chen, E., Daumke, O., Faelber, K., Ford, M., Frolov, A.V., Frost, A., Hinshaw, J.E., Kirchhausen, T., Kozlov, M.M., Lenz, M., Low, H.H., McMahon, H., Merrifield, C., Pollard, T.D., Robinson, P.J., Roux, A., & Schmid, A. (2016). Membrane fission by dynamin: what we know and what we need to know. The EMBO Journal, 35(21), 2270-2284.
Buj-Bello, A., Laugel, V., Messaddeq, N., Zahreddine, H., Laporte, J., Pellissier, K.F., et al. (2002). The lipid phosphatase myotubularin is essential for skeletal muscle maintenance but not for myogenesis in mice. Ophanet Journal of Rare Diseases, 3(5), 35. Chin, Y.-H., Lee, A., Kan, H.-W., Laiman, J., Chuang, M.-C., Hsieh, A.-T., & Liu, Y.-W. (2015). Dynamin-2 mutations associated with centrouclear myopathy are hypermorphic and lead to T-tubule fragmentation. Human Molecular Genetics, 24, 5542-5554. Cowling, B.S., Toussaint, A., Amoasii, L., Koebel, P., Ferry, A., Davignon, L., Nishino, I., Mandel, J.L., & Laporte, J. (2011). Increased expression of wild-type or a centronuclear myopathy mutant of dynamin 2 in skeletal muscle of adult mice leads to structural defects and muscle weakness. The American Journal of Pathology, 178, 2224-2235. Cowling, B.S., Chevremont, T., Prokic, I., Kretz, C., Ferry, A., Coirault, C., Koutsopoulos, O., Laugel, V., Romero, N.B., & Laporte, J. (2014). Reducing dynamin 2 expression X-linked centronuclear myopathy. Journal of Clinical Investigation, 124(3), 1350-1363. Cowling, B.S., Prokic, I., & Tasfaout, H., Rabai, A., Humbert, F., Rinaldi, B., Nicot, A., Kretz, C., Friant, S., Roux, A., & Laporte, J. (2017). Amphiphysin (BIN1) negatively regulates dynamin 2 for normal muscle maturation. The Journal of Clinical Investigation, 127(12), 4477-4487. Daumke, O., Roux, A., & R., Haucke, V. (2014). BAR domain scaffolds in dynamin-mediated membrane fission. Cell, 156, 882-892. Demonbreun, A.R., & McNally, E.M. (2014). Dynamin 2 the rescue for centronuclear myopathy. Journal of Clinical Investigation, 124(3), 976-978. Gibbs, E.M., Davidson, A.E., Telfer, W.R., Feldman, E.L., & Dowling, J.J. (2014). The myopathy-causing mutation DNM2-S619L leads to defective tubulation in vitro and in developing zebrafish. Disease Models and Mechanisms, 7, 157-161. Hohendahl, A., Roux, A., & Galli, V. (2016). Structural insights into the centronuclear myopathy-associated functions of BIN1 and dynamin 2. Journal of Structural Biology, 196, 37-47. Hohendahl, A., Talledge, N., Galli, V., Shen, P.S., Humbert, F., De Camilli, P., Frost, A., & Roux, A. (2017). Structural inhibition of dynamin-mediated membrane fission by endophilin. Elife, 6, pii: e26856. Kenniston, J.A., & Lemmon, M.A. (2010). Dynamin GTPase regulation is altered by PH domain mutations found in centronuclear myopathy patients. EMBO Journal, 29, 3054-3067. Kojima, C., Hashimoto, A., Yabuta, I., Hirose, M., Hashimoto, S., Kanaho, Y., Sumimoto, H., Ikegami, T., & Sabe, H. (2004). Regulation of Bin1 SH3 domain binding by phosphoinositides. EMBO Journal, 23, 219-228. Lee, E., Marucci, M., Daniell, L., Pypaert, M., Weisz, O.A., Ochoa, G., Farsad, K., Wenk, M.R., & Camilli, P.D. (2002). Amphiphysin 2 (Bin1) and T-tubule biogenesis in muscle. Science, 297, 1193. Liu, Y.-W., Lukiyanchuk, V., & Schmid, S.L. (2011). Common membrane trafficking defects of disease-associated dynamin 2 mutations. Traffic, 12, 1620-1633. Meinecke, M., Boucrot, E., Camdere, G., Hon, W., Mittal, R., & McMahon, H.T. (2013). Cooperative recruitment of dynamin and BIN/amphiphysin/Rvs (BAR) domain-containing proteins leads to GTP-dependent membrane scission. The Journal of Biological Chemistry, 288(9), 6651-6661. Neumann, S., & Schmid, S.L. (2013). Dual role of BAR domain-containing proteins in regulating vesicle release catalyzed by the GTPase, dynamin-2. The Journal of Biological Chemistry, 288(35), 2519-25128. Nicot, A., Toussaint, A., Tosch, V., Kretz, C., Wallgren-Pettersson, C., Iwarsson, E., Kingston, H., Garnier, J., Biancalana, V., Oldfors, A., Mandel, J., & Laporte, J. (2007). Mutations in amphiphysin (BIN1) disrupt interaction with dynamin 2 and cause autosomal recessive centronuclear myopathy. Nature Genetics, 39(9), 1134-1139. Owen, D.J., Wigge, P., Vallis, Y., Moore, J.D.A., Evans, P.R., & McMahon, H.T. (1998). Crystal structure of the amphiphysin-2 SH3 domain and its role in the prevention of dynamin ring formation. The EMBO Journal, 17(18), 5273-5285. Prokic, I., Cowling, B.S., & Laporte, J. (2014). Amphiphysin 2 (BIN1) in physiology and diseases. Journal of Molecular Medicine, 92, 453-463. Syndborger, A., Soderblom, C., Vorontsova, O., Evergren, E., Hinshaw, J.E., & Shupliakov, O. (2011). An endophilin-dynamin complex promotes budding of clathrin-coated vesicles during synaptic vesicle recycling. Journal of Cell Science, 124(1), 133-143. Wu, T., Shi, Z., & Baumgart, T. (2014). Mutations in BIN1 associated with centronuclear myopathy disrupt membrane remodeling by affecting protein density and oligomerization. PLoS One, 9(4), e93060. Antonescu, C.N., Danuser, G. & Schmid, S.L. (2010). Phosphatidic acid plays a regulatory role in clathrin-mediated endocytosis. Molecular and Cellular Biology, 21(16), 2944-2952. Bruntz, R.C., Lindsley, C.W., & Brown, H.A. (2014). Phosphlipase D signaling pathways and phosphatidic acid as therapeutic targets in cancer. Pharmacological Reviews, 66(4), 1033-1079. Diz-Munoz, A., Fletcher, D.A., & Weiner, O.D. (2013). Use the force: Membrane tension as an organizer of cell shape and motility. Trends in Cell Biology, 23(2), 47-53. Hornberger, T.A., Chu, W.K., Mak, Y.W., Hsiung, J.W., Huang, S.A., & Chien, S. (2006). The role of phospholipase D and phosphatidic acid in the mechanical activation of mTOR signaling in skeletal muscle. Proceedings of the National Academy of Sciences of the United States of America, 103(12), 4741-4746. Lim, J.P., & Gleeson, P.A. (2011). Macropinocytosis: an endocytic pathway for internalising large gulps. Immunology and Cell Biology, 89, 836-843. Pelkmans, L., Puntener, D., & Helenius, A. (2002). Local actin polymerization and dynamin recruitment in SV40-induced internalization of caveolae. Science, 296(5567), 535-539. Petersen, E.N., Chung, H.-W., Nayebosadri, A., & Hansen, S.B. (2016). Kinetic disruption of lipid rafts is a mechanosensory for phospholipase D. Nature Communications, 7. Pontes, B., Monzo, P., & Gauthier, N.C. (2017). Membrane tension: A challenging but universal parameter in cell biology. Seminars in Cell & Developmental Biology, 71, 30-41. Wu, X.S., Elias, S., Liu, H., Heureaux, J., Wen, P.J., Liu, A.P., Kozlov, M.M., & Wu, L.G. (2017). Membrane tension inhibits rapid and slow endocytosis in secretory cells. Biophysical Journal, 113(11), 2406-2414. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/69569 | - |
| dc.description.abstract | 中央核肌肉病變 (Centronuclear myopathy, CNM)是一種罕見的遺傳疾病，病徵是骨骼肌的衰弱與退化。CNM分為三種類型，其中的體染色體隱性遺傳型是由一個骨骼肌中負責橫小管(T-tubule)發育的蛋白Bin1突變所造成。T-tubule是骨骼肌細胞膜內凹陷入細胞內的結構，對肌肉細胞的興奮收縮連結作用(E-C coupling)非常重要。許多造成CNM的Bin1突變體是發生在產生膜彎曲的區域進而產生T-tubule發生的缺陷。然而，有兩個造成Bin1 SH3區域截斷的無義突變(nonsense mutations)體， Q434X與K436X，目前仍不清楚其致病機制。在這篇論文中，我發現這兩個Bin1突變體喪失了對Dynamin 2(Dyn2)的調控而造成T-tubule無法維持。Dyn2是一個全身性表達的細胞膜切斷酵素。從體外實驗的結果，我發現Bin1 Q434X與K436X 對Dyn2的結合力下降因此造成Dyn2切斷膜的能力上升。相同的影響也在肌纖維母細胞中利用共同表達Bin1 Q434X-GFP 或K436X-GFP 與 Dyn2-mCherry 觀察到。因此，我們發現Bin1是Dyn2活性的調控蛋白，而Bin1 Q434X與K436X突變造成了Dyn2的過度活躍而造成T-tubule斷裂而產生了骨骼肌的病變。我的實驗結果除了提供CNM的致病機制，並指出降低Dyn2的活性將是治療Bin1-CNM的一個方法。 | zh_TW |
| dc.description.abstract | Centronuclear myopathy (CNM) is a rare genetic disorder characterized by muscle weakness that is debilitating from an early age. Autosomal recessive CNM is caused by mutations in Amphiphysin-2 (Bin1), a protein important for membrane curvature generation and critical for the formation of highly curved plasma membrane invaginations in muscle cells, the T-tubules, which are crucial for excitation-contraction coupling. There have been several CNM-related Bin1 mutations found to be located at its membrane curvature generating domain or membrane binding domain resulting in defects in Bin1’s tubulation ability. Two nonsense mutations, Q434X and K436X, which result in truncated Src homology 3 (SH3) domains, have not been well-characterized for their pathogenic mechanism. The phenotypes of these Bin1 mutants are illustrated in relation to their effects on dynamin-2 (Dyn2) activity in this study. Dyn2 is a GTPase that is ubiquitously expressed that is well-known for its scission of vesicles from the membrane. I discovered that Bin1 mutants, Q434X and K436X, have an enhancing effect on Dyn2 fission activity because Bin1 Q434X and K436X are diminished in their binding affinity with Dyn2 compared to full-length Bin1. Consistent results were also observed in vivo by co-transfection of the Q434X-GFP or K436X-GFP with Dyn2-mCherry in C2C12 myoblast cells. In conclusion, wild-type Bin1 was found to negatively regulate dynamin fission activity and the hyperactivity quality of Dyn2 membrane fission underpins the autosomal recessive CNM caused by SH3 domain truncated Bin1. Together, my study provides a mechanistic explanation for the characteristics of T-tubule fragmentation displayed by CNM and implicates downregulation of dynamin to be a therapeutic strategy in mitigating muscle atrophy by CNM. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-17T03:19:37Z (GMT). No. of bitstreams: 1 ntu-107-R05448016-1.pdf: 3729731 bytes, checksum: 718701da3f14bb9c1ed201e52e3f310c (MD5) Previous issue date: 2018 | en |
| dc.description.tableofcontents | Table of Contents
論文審定書 i Acknowledgement ii 中文摘要 iii ABSTRACT iv Chapter 1 INTRODUCTION 1 1.1 Forms of Centronuclear Myopathy 1 1.2 Bin1 Sculpts the T-Tubule Membrane 2 1.3 Dynamin and Regulation by Bin1 3 1.4 CNM-linked Mutations in H0 domain, BAR domain, and PI motif of Bin1 4 1.5 CNM-linked Mutations in SH3 domain of Bin1 5 Chapter 2 MATERIALS AND METHODS 7 2.1 Bin1 Cloning and Protein Purification 7 2.2 Q434X and K436X Point Mutagenesis, Cloning, and Proteins 7 2.3 Generation of SUPER templates 8 2.4 Sedimentation (Fission) Assay 8 2.5 Tubulation Assay 8 2.6 Pulldown Assay with Glutathione-S-Transferase (GST) PRD of Dyn2 9 2.7 Liposome Binding Assay 9 2.8 Transmission Electron Microscopy 10 2.9 Transfection of C2C12 cells and Fixed Imaging 10 2.10 Blind Test Quantification of Phenotypes of C2C12 myoblast cells 10 2.11 Live Cell Imaging of Transfected C2C12 myoblast cells 11 Chapter 3 RESULTS 12 3.1 Bin1 Tubulates the Membrane in vitro 12 3.2 Dynamin Interferes with Bin1 Membrane Tubulation in vitro 12 3.3 Decreased Binding of Q434X and K436X to PRD of Dyn2 13 3.4 Bin1 and Dyn2 Binding to Membrane 14 3.5 Bin1 and Dyn2 Assembly on Membrane under TEM 15 3.6 Bin1 and Dyn2 Assembly Together on Membrane under TEM 16 3.7 Bin1 Negatively Regulates Dynamin in vitro 17 3.8 Q434X and K436X Enhance Dynamin Fission in vitro 18 3.9 Bin1 Tubulates the Membrane in vivo 19 3.10 Vesicle Formation in Co-Transfection of Mutants with Dyn2 19 3.11 Dynamic Fission Events in vivo under Live Cell Imaging 20 Chapter 4 DISCUSSION 21 4.1 Bin1 Negatively Regulates Dynamin 21 4.2 Dyn2 Hyperactivity with Q434X and K436X 22 4.3 Dynamin as a Therapeutic Target for CNM 23 Chapter 5 FIGURES 26 Figure 1 Domains of muscle-specific Amphiphysin-II (Bin1) 26 Figure 2 Membrane tubulation by WT Bin1, Q434X, and K436X 28 Figure 3 Membrane tubulation by WT Bin1, Q434X, and K436X with dynamin 30 Figure 4 Population of SUPER template beads with tubules 32 Figure 5 Pulldown of WT Bin1, Q434X, and K436X by GST-PRD of Dyn2 33 Figure 6 Liposome binding of Dyn2, WT Bin1, Q434X and K436X 36 Figure 7 Assembly of Bin1 and Dyn2 under TEM 38 Figure 8 Assembly of Bin1 with Dyn2 together under TEM 40 Figure 9 Bin1 and Dynamin fission activity 41 Figure 10 Bin1-SH3 and Dyn1 fission activity 44 Figure 11 C2C12 myoblast cells transfected with Bin1-GFP 46 Figure 12 C2C12 cells co-transfected with Dyn2-mCherry and Bin-GFP 48 Figure 13 Live cell imaging of co-transfected C2C12 myoblast cells 50 Figure 14 Pathogenic mechanism of BIN1 SH3-CNM 51 Chapter 6 REFERENCES 54 APPENDIX: Membrane tension decrease induces macropinocytosis through PLD activity 56 ABSTRACT 56 I INTRODUCTION 57 I.I Plasma Membrane Tension Sensing and Feedback 57 I.II Potential for Lipid Rafts as Signaling Platforms 58 II MATERIALS AND METHODS 60 II.I C2C12 Myoblast Cells and Myotube Cells 60 II.II Phosphatidic Acid Assay 60 II.III Dynamin GTP Hydrolysis 61 II.IV Sedimentation (Fission) Assay 61 II.V Liposome Binding Assay 61 II.VI Transmission Electron Microscopy 62 III RESULTS 63 III.I Phosphatidic Acid Accumulates When Membrane Tension Changes 63 III.II Phospholipase D Induces PA Production 64 III.III Dynamin Supports Membrane Tension Homeostasis Through PA 66 IV DISCUSSION 68 IV.I Lipid Rafts Function as Signaling Platforms Containing PLD2 68 IV.II PLD2 Activates Macropinocytosis When Membrane Tension Plunges 69 V FIGURES 72 Figure 1 Distribution of PABD-GFP upon changes in membrane tension 72 Figure 2 Phosphatidic acid content upon changes in membrane tension 74 Figure 3 Phosphatidic acid content with PLD inhibitors 76 Figure 4 Macropinocytosis in C2C12 myoblast cells with PLD inhibitors 78 Figure 5 Macropinocytosis in C2C12 myotube cells 80 Figure 6 Distribution of PA, PLD1, and PLD2 in macropinosomes in C2C12 myotube cells 82 Figure 7 Crowding of macropinosomes in C2C12 myotube cells 84 Figure 8 Localization of dynamin and actin near membrane ruffles in C2C12 myoblast cells 86 Figure 9 Dynamin GTP hydrolysis with increasing concentrations of PA 88 Figure 10 Dynamin fission activity with increasing concentrations of PA 90 Figure 11 Tubulation of dynamin with increasing concentrations of PA 91 Figure 12 Liposome binding dynamin with increasing concentrations of PA 93 Figure 13 Assembly of dynamin with increasing concentrations of PA 95 VI REFERENCES 98 | |
| dc.language.iso | en | |
| dc.subject | 膜販運 | zh_TW |
| dc.subject | 體染色體隱性遺傳中央核肌肉病變 | zh_TW |
| dc.subject | 膜張力 | zh_TW |
| dc.subject | 肌肉細胞 | zh_TW |
| dc.subject | 致病機制 | zh_TW |
| dc.subject | 肌肉病 | zh_TW |
| dc.subject | centronuclear myopathy | en |
| dc.subject | dynamin | en |
| dc.subject | Amphiphysin-2 | en |
| dc.subject | Bin1 | en |
| dc.subject | membrane trafficking | en |
| dc.subject | membrane remodeling | en |
| dc.subject | membrane fission | en |
| dc.title | 體染色體隱性遺傳中央核肌肉病變的致病機制 | zh_TW |
| dc.title | Pathogenic Mechanism of Autosomal Recessive Centronuclear Myopathy | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 106-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 蔡丰喬(Feng-Chiao Tsai),陳文彬(Wen-Pin Cheng),蔡欣祐(Hsin-Yue Tsai) | |
| dc.subject.keyword | 肌肉病,致病機制,肌肉細胞,膜張力,膜販運,體染色體隱性遺傳中央核肌肉病變, | zh_TW |
| dc.subject.keyword | centronuclear myopathy,dynamin,Amphiphysin-2,Bin1,membrane trafficking,membrane remodeling,membrane fission, | en |
| dc.relation.page | 106 | |
| dc.identifier.doi | 10.6342/NTU201800973 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2018-06-27 | |
| 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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