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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 鄭萬興(Wan-Hsing Cheng) | |
| dc.contributor.author | Wei-Chih Lin | en |
| dc.contributor.author | 林威至 | zh_TW |
| dc.date.accessioned | 2022-11-23T08:57:37Z | - |
| dc.date.available | 2022-01-17 | |
| dc.date.available | 2022-11-23T08:57:37Z | - |
| dc.date.copyright | 2022-01-17 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-11-22 | |
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(2013) Systematic exploration of thioredoxin target proteins in plant mitochondria. Plant Cell Physiol. 54: 875-892. Zimorski, V., Ku, C., Martin, W.F. and Gould, S.B. (2014) Endosymbiotic theory for organelle origins. Curr. Opin. Microbiol. 22: 38-48. Zmudjak, M., Colas des Francs‐Small, C., Keren, I., Shaya, F., Belausov, E., Small, I. and Ostersetzer‐Biran, O. (2013) m CSF 1, a nucleus‐encoded CRM protein required for the processing of many mitochondrial introns, is involved in the biogenesis of respiratory complexes I and IV in Arabidopsis. New Phytol. 199: 379-394. Zmudjak, M., Shevtsov, S., Sultan, L.D., Keren, I. and Ostersetzer-Biran, O. (2017) Analysis of the roles of the Arabidopsis nMAT2 and PMH2 proteins provided with new Insights into the regulation of Group II intron splicing in land-plant mitochondria. Int. J. Mol. Sci. 18: 2428. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/79296 | - |
| dc.description.abstract | 粒線體是由原核細菌(α-proteobacteria)演化而來，在長期互利共生的關係下，逐漸演化成真核細胞的能量工廠並轉移了多數的基因到核基因組(nuclear genome)。因此，維持粒線體的呼吸作用正常運作並產生能量，需要核基因所編碼的蛋白質對粒線體基因進行多種後轉錄修飾。特別是許多組成粒線體呼吸傳遞鏈的蛋白質，這些編碼蛋白質的基因帶有退化的Group II 內含子，己失去自行剪切的能力，需要協同多種細胞核的蛋白質來完成內含子剪切(intron splicing)。CRM蛋白家族是細胞核編碼的核酸結合蛋白，目前多數的研究報導CRM蛋白經由參與group I 和group II內含子剪切來調控葉綠體基因的表現。然而CRM蛋白在粒線體中扮演的生理功能及作用機制仍未完全明瞭。本研究進一步分析阿拉伯芥CFM6基因的生理功能及分子機制，並揭開這個具單一CRM蛋白結構域(CRM domain)的家族成員在粒線體中扮演的角色。當T-DNA插入造成CFM6基因突變，會嚴重影響cfm6突變株的發育，包含生長遲緩、葉片捲曲、種子皺縮、延遲胚及花粉的發育。實驗結果顯示CFM6蛋白具專一性調控粒線體nad5基因第四個內含子的剪切；因此，在cfm6突變株中造成未經剪切的轉錄物(pretranscripts)。於cfm6突變株中大量表現CFM6-YFP（CFM6蛋白融合黃色螢光蛋白）可恢復植株發育不良的性狀及內含子剪切功能的缺失。因nad5蛋白為組成呼吸傳遞鏈複合體 I (respiratory complex I) 的元件，其轉錄後內含子剪切的缺失進一步造成cfm6突變株粒線體內複合體 I 的生合成降低及活性損壞，伴隨粒線體形態發育異常，並且誘導替代呼吸途徑(alternative respiration pathway)相關基因的表現。根據RNA-seq分析的結果，CFM6基因突變擾動細胞核及胞器基因之表現，全數2764個受擾動的基因中，多數與光合作用相關的基因表量下降，而與核糖體生合成相關的基因表現量則呈現上升的趨勢。目前的研究結果顯示CFM6是一個參與粒線體內nad5基因第四個內含子剪切的剪切因子，在粒線體呼吸作用複合體 I 的生合成及植物生長發育過程中扮演重要的角色。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-23T08:57:37Z (GMT). No. of bitstreams: 1 U0001-1611202116042600.pdf: 16242565 bytes, checksum: 684481a8b857fa7668b166a7ab7eb198 (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | 英文摘要 ....V List of figures and tables ....IX 1. Introduction ....1 1.1 Endosymbiotic origin of mitochondria ....1 1.2 The role of mitochondria ....1 1.3 Mitochondrial gene expression ....4 1.4 Group II intron splicing ....5 1.5 Splicing factors ....6 1.6 CRM domain proteins ....8 1.7 Aim of this study ....9 2. Materials and Methods ....10 2.1 Plant materials and growth conditions ....10 2.2 Complementation test ....11 2.3 RT-PCR and RT-qPCR analyses ....11 2.4 Blue native gel electrophoresis (BN-PAGE) and immunoblot detection ....12 2.5 Microscopy analyses ....13 2.6 Statistical analyses ....13 2.7 RNA-seq and gene ontology (GO) analysis ....14 2.8 Yeast two hybrid (Y2H) interaction assay ....15 3. Results ....16 3.1 Mutation of CFM6 causes defects in the development of vegetative and reproductive tissues in Arabidopsis cfm6 mutant ....16 3.2 Degenerative pollen grains occur predominantly at the mature stage in cfm6 mutant ....18 3.3 complementation and functional rescue test of cfm6 mutant ....20 3.4 Tissue-specific expression and subcellular localization of CFM6 ....21 3.5 CFM6 is required for cis-splicing of mitochondrial nad5 intron 4 ....22 3.6 CFM6 affects mitochondrial complex I function and biogenesis ....25 3.7 Mitochondria of cfm6 have an altered structure and are less sensitive to the complex I inhibitor rotenone ....28 3.8 CFM6-mediated gene expression profiling ....30 4. Discussion ....31 4.1 CFM6 functions in the specific splicing of the nad5 intron 4 and affects mitochondrial complex I biogenesis and normal plant growth ....31 4.2 Male sterility occurs at the late developmental stage of gametophytes in cfm6 mutants ....35 5. Conclusions and outlook ....38 References ....40 Appendix ....69 Figure S1 ....69 Figure S2 ....70 Figure S3 ....71 Figure S4 ....72 Preparations for the crude mitochondria isolation and complex I activity assay ....73 | |
| dc.language.iso | en | |
| dc.title | 阿拉伯芥細胞核編碼的一個粒線體CRM結構域蛋白CFM6之功能性分析 | zh_TW |
| dc.title | "Functional characterization of CFM6, a nuclear-encoded mitochondrial CRM domain-containing protein, in Arabidopsis thaliana" | en |
| dc.date.schoolyear | 110-1 | |
| dc.description.degree | 博士 | |
| dc.contributor.author-orcid | 0000-0003-3262-2755 | |
| dc.contributor.advisor-orcid | 鄭萬興(0000-0003-4924-7429) | |
| dc.contributor.coadvisor | 張英峯(Ing-Feng Chang) | |
| dc.contributor.oralexamcommittee | 洪傳揚(Hsin-Tsai Liu),張孟基(Chih-Yang Tseng),趙光裕 | |
| dc.subject.keyword | 阿拉伯芥,CRM蛋白結構域,胚胎發生,Group II內含子,雄不稔,粒線體NADH去氫酶, | zh_TW |
| dc.subject.keyword | Arabidopsis thaliana,CRM domain-containing protein,Embryogenesis,Group II intron,Male sterility,Mitochondrial respiratory complex I, | en |
| dc.relation.page | 76 | |
| dc.identifier.doi | 10.6342/NTU202104481 | |
| dc.rights.note | 同意授權(全球公開) | |
| dc.date.accepted | 2021-11-22 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 植物科學研究所 | zh_TW |
| 顯示於系所單位： | 植物科學研究所 | |
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