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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 謝旭亮 | |
dc.contributor.author | Yi-Hung Tsai | en |
dc.contributor.author | 蔡易宏 | zh_TW |
dc.date.accessioned | 2021-06-15T12:27:46Z | - |
dc.date.available | 2018-10-14 | |
dc.date.copyright | 2016-10-14 | |
dc.date.issued | 2016 | |
dc.date.submitted | 2016-08-09 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/50013 | - |
dc.description.abstract | 植物生長發育過程受數種荷爾蒙及多項環境因子如光等共同調控。幼苗發育早期,活化的遠紅光受體光敏素A(phytochrome A)、紅光受體光敏素B(phytochrome B)與藍光受體隱花色素(cryptochrome)皆參與荷爾蒙相關信息傳遞。近年研究指出FAR-RED INSENSITIVE 219 (FIN219)具催化茉莉酸(jasmonate)與異白胺酸(isoleucine)結合並生成具生物活性的茉莉酸外,更涉及藍光及遠紅光等傳訊路徑。實驗團隊先前亦發現受酪蛋白激酶(Casein kinase 2)所調控之FIN219蛋白質磷酸化將使其酵素活性改變。本研究嘗試透過FIN219蛋白質受酪蛋白激酶磷酸化位點突變的轉殖株,釐清其磷酸化狀態於植物生長發育早期所扮演的角色,並藉磷酸標籤蛋白質電泳,尋找可能利用FIN219為受質之磷酸化激酶。初步結果顯示,FIN219蛋白質兩個受酪蛋白激酶2磷酸化的胺基酸突變時,將造成FIN219蛋白質於藍光照射下穩定性改變,且此兩個胺基酸改變的雙突變體幼苗下胚軸長度於藍光下亦明顯低於野生型。另磷酸標籤蛋白電泳顯示,在黑暗、遠紅光與紅光中生長的野生型幼苗,含有磷酸化的FIN219蛋白質;相對的遠紅光及紅光照射下對野生型幼苗外源添加甲基茉莉酸,以及遠紅光下光敏素A突變體、黑暗中光敏素B突變體及藍光下酪蛋白激酶突變體皆造成幼苗累積較多未磷酸化的FIN219蛋白質。反之短暫藍光照射則可能藉隱花色素相關路徑,造成FIN219蛋白質去磷酸化。同時,體外磷酸激酶試驗亦指出光敏素A與光敏素B皆可造成FIN219蛋白質磷酸化,且未磷酸化之FIN219蛋白質呈現與PHYA及COP1蛋白質更高的親和力。綜合上述,光敏素及隱花色素皆可能透過調控FIN219蛋白質磷酸化狀態,以調控光型態發生的反應 。 | zh_TW |
dc.description.abstract | Several hormones and photoreceptors together regulate photomorphogenesis of seedlings. To respond to various environments, post-translational modifications such as kinase-mediated phosphorylation have been demonstrated to participate in several light signaling pathways. Previous studies revealed that FIN219/JAR1, a jasmonate conjugating enzyme, could be phosphorylated by casein kinase 2 (CK2). Enzyme kinetic analysis also showed that unphosphorylated FIN219 exhibited higher substrate affinity. To further determine the roles of two phosphorylation isoforms of FIN219, transgenic lines containing FIN219 native promoter driven substitution of residues that were demonstrated to be phosphorylated by CK2 showed the phenotypic effects on methyl jasmonate (MeJA)-mediated inhibition of hypocotyl elongation and increased accumulations of dephosphorylated FIN219 under blue light. On the other hand, Phos-tagTM acrylamide SDS-PAGE electrophoresis showed that most of FIN219 would be phosphorylated in the dark, red and far-red light, but exogenous MeJA could trigger dephosphorylation of FIN219 under far-red light and red light. Additionally, kinase assays indicated that PHYA and PHYB could phosphorylate FIN219 in vitro, which is consistent with the fact that only the unphosphorylated form of FIN219 exists in both phyA null mutant under far-red light and phyB null mutant in the dark. Protein interaction assays also revealed that dephosphorylated FIN219 exhibited higher affinity
to PHYA and COP1. Interestingly, blue light irradiation resulted in dephosphorylation of FIN219 in a cryptochromes-dependent manner. Taken together, current data indicate that photoreceptor-mediated phosphorylation changes of FIN219 plays a vital role in regulating photomorphogenic development of Arabidopsis seedlings. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T12:27:46Z (GMT). No. of bitstreams: 1 ntu-105-R03b42004-1.pdf: 2665200 bytes, checksum: 041c84e1f7ee4156c0989c03dc2e00fb (MD5) Previous issue date: 2016 | en |
dc.description.tableofcontents | 摘要 IV
Abstract IV 前言 1 一. 緒論 1 二. 光敏素調控的光訊息傳遞 2 三. 光敏素A的磷酸化激酶活性 3 四. 酪蛋白激酶(Casein kinase 2, CK2)的生理功能 4 五. 茉莉酸的合成與訊息傳導 4 六. FIN219/JAR1為遠紅光及茉莉酸訊號傳遞的整合蛋白質 5 七. 隱花色素CRY1,CRY2與FIN219間的調控關係 6 研究目標 7 材料與方法 8 一. 植物材料與生長條件 8 二. 體外磷酸化試驗分析 8 三. 融合蛋白質的誘導表現與純化 8 四. 磷酸標籤(Phos-tagTM)蛋白質電泳 9 五. 植物蛋白質表現量分析 9 六. RNA萃取與RNA表現分析 9 七. 蛋白酶體活性抑制劑MG132處理 10 八. 蛋白質共免疫沉澱分析 10 九. Pull-down assay 10 結果 11 一. 外源添加甲基茉莉酸將造成植物體中FIN219蛋白質去磷酸化 11 二. 光敏素A與光敏素B融合蛋白質可磷酸化FIN219蛋白質 11 三. 光敏素A為遠紅光下FIN219蛋白質磷酸化的主要激酶 12 四. FIN219磷酸化狀態會影響與COP1及光敏素A的交互作用 13 五. 酪蛋白激酶CK2為藍光下FIN219蛋白質磷酸化的主要激酶 14 六. 去磷酸化的FIN219蛋白質可受26S蛋白酶體相關路徑所分解 14 七. 短暫藍光照射造成FIN219蛋白質去磷酸化 15 八. FIN219蛋白質磷酸化位點突變轉殖株呈現藍光不敏感表型 15 九. 藍光下FIN219蛋白質第55號及69號蘇胺酸磷酸化影響其蛋白質穩定性 16 十. 數種光接受體參與FIN219蛋白質的磷酸化 17 一. 茉莉酸對FIN219蛋白質磷酸化的影響 19 二. 光敏素所調控的FIN219蛋白質磷酸化 20 三. FIN219蛋白質與COP1及光敏素A之交互作用 22 四. 藍光中未知的去磷酸化酶與酪蛋白激酶共同調控FIN219蛋白質的磷酸化狀態 23 五. FIN219蛋白質的磷酸化與光型態發生 24 結論 26 結果圖片 27 參考文獻 38 附錄一、實驗操作流程 49 附綠二、PCR相關資訊 58 圖目錄 圖一. 外源添加甲基茉莉酸造成FIN219蛋白質去磷酸化 27 圖二. 光敏素及酪蛋白激酶皆可於體外磷酸化FIN219蛋白質 29 圖三. 光敏素A為遠紅光下FIN219蛋白質磷酸化的主要激酶 29 圖四. FIN219蛋白質磷酸化狀態可影響與COP1蛋白質及光敏素A蛋白質的交互作用 30 圖五. 去磷酸化的FIN219蛋白質可受26S蛋白酶體相關路徑所分解 31 圖六. 酪蛋白激酶CK2為藍光下FIN219蛋白質磷酸化的主要激酶 32 圖七. FIN219蛋白質受CK2磷酸化位點突變轉植株呈現藍光不敏感表型 34 圖八. FIN219蛋白質第55號及69號蘇胺酸磷酸化影響藍光下其蛋白質穩定性 35 圖九. 數種光接受體參與FIN219蛋白質的磷酸化 36 圖十. FIN219蛋白質磷酸化後轉譯修飾與光訊息傳遞路徑的關係 37 | |
dc.language.iso | zh-TW | |
dc.title | 阿拉伯芥中FIN219/JAR1蛋白質磷酸化修飾在光與茉莉酸訊息傳遞中的功能性研究 | zh_TW |
dc.title | Functional studies of FIN219/JAR1 phosphorylation involved in integration of light and jasmonate signaling in Arabidopsis | en |
dc.type | Thesis | |
dc.date.schoolyear | 104-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 鄭萬興,吳素幸,涂世隆,吳克強 | |
dc.subject.keyword | FIN219,磷酸化修飾,茉莉酸,光型態發生,酪蛋白激?, | zh_TW |
dc.subject.keyword | FIN219,phosphorylation,jasmonate,photomorphogenesis,casein kinase 2, | en |
dc.relation.page | 58 | |
dc.identifier.doi | 10.6342/NTU201602093 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2016-08-09 | |
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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