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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 吳益群 | |
dc.contributor.author | Meng-I Lee | en |
dc.contributor.author | 李孟怡 | zh_TW |
dc.date.accessioned | 2021-06-16T13:20:59Z | - |
dc.date.available | 2018-08-23 | |
dc.date.copyright | 2013-08-23 | |
dc.date.issued | 2013 | |
dc.date.submitted | 2013-07-25 | |
dc.identifier.citation | Calixto, Andrea; Chelur, Dattananda; Topalidou, Irini; Chen, Xiaoyin; Chalfie, Martin (2010). Enhanced neuronal RNAi in C. elegans using SID-1. Nature methods. 7(7):554-9
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/61968 | - |
dc.description.abstract | 計畫性細胞死亡的主要功能是在維持體內細胞數的平衡以及移除掉不想要的細胞,如果計畫性細胞死亡的調控過程中發生問題可能會造成癌症以及某些疾病。在過去30年中,已知有數十個基因,包括主要進行細胞死亡的基因: EGL-1(a BH3-containing protein),CED-9(Bcl-2),CED-4(Apaf-1)以及CED-3(Caspase)已經被知道參與在線蟲計畫性細胞死亡中執行死亡的階段,然而這4個主要的基因是如何被調控的以及還有什麼樣的基因參與在其中還不是非常清楚。在這篇論文中,我發現鈉鉀ATP酶α單元EAT-6可以促進線蟲的計畫性細胞死亡。哺乳動物的鈉鉀ATP酶由2個含有ATP酶活性的α單元和2個β單元以及1個在特定組織中才有的γ單元所組成,它主要的功能是在維持細胞膜電位的平衡。而線蟲有2個α單元eat-6和catp-4和3個β單元nkb-1,nkb-2和nkb-3,但是沒有γ單元。在這裡我發現失去任何的β單元不會造成胚胎發育過程中計畫性細胞死亡的問題,但是一旦失去α單元eat-6就會造成胚胎發育過程中應該要死的細胞沒死掉,這個結果顯示α單元與β單元參與在計畫性細胞死亡的角色並不相同。除此之外,當大量表現eat-6時會促進細胞死亡。很有趣的是,當EAT-6的ATP酶活性位置如果發生突變的話,仍然會促進細胞死亡,因此代表EAT-6除了在運送鈉鉀離子的功能之外還有另一個新的角色,也就是促進細胞死亡。另外在特定細胞中,沒有結合β單元NKB-1或 NKB-2的α單元EAT-6也會促進細胞死亡,代表在這些細胞中單獨α單元EAT-6就有促進死亡的功能了。除了eat-6之外,我還有發現蛋白激酶A (cAMP-dependent protein kinase)的調控單元kin-2也會促進線蟲的計畫性細胞死亡。在哺乳動物中,蛋白激酶A是由2個調控單元以及2個活化單元所組成,但是線蟲只有1個調控單元kin-2和 1個活化單元kin-1。在這裡我發現,一旦失去調控單元kin-2就會造成胚胎發育過程中應該要死的細胞沒死掉,但是當失去活化單元kin-1反而會更增加細胞的死亡。除此之外,當大量表現kin-2時也會促進細胞死亡。然而我還發現在dying cell中表現eat-6 或kin-2會挽救eat-6 或kin-2突變蟲減少細胞死亡的問題,因此顯示eat-6 和kin-2是會作用在dying cell的。另外eat-6和kin-2是會透過egl-1幫助去促進計畫性細胞死亡的。 | zh_TW |
dc.description.abstract | Programmed cell death (apoptosis) functions in maintaining homeostasis and eliminating unwanted cells. Defects in programmed cell death has been implicated in cancerous growth and diseases. Over the past 30 years, about a dozen of genes, including the core cell death genes EGL-1 (a BH3-containing protein), CED-9 (Bcl-2), CED-4 (Apaf-1) and CED-3 (Caspase), have been identified to function in the execution of programmed cell death in C. elegans. However, how the cell death process is regulated is still unclear. Here, I report that EAT-6, a sodium-potassium ATPase alpha subunit, can promote programmed cell death in C. elegans. A typical mammalian sodium-potassium ATPase is composed of two alpha subunits containing an ATPase activity and two beta subunits with/without one gamma subunit, and functions to maintain membrane potential across plasma membrane. C. elegans has two alpha subunits, eat-6 and catp-4, and three beta subunits, nkb-1, nkb-2 and nkb-3, but has no gamma subunit. I found that loss of any beta subunit does not result in a detectable cell death defect during embryogenesis and, in contrast, loss of the alpha subunit eat-6, significantly reduced cell death during embryogenesis. This result shows differential involvement of alpha and beta subunits in promoting cell death. In addition, overexpression of eat-6 promotes cell death. Interestingly, the “pump dead” EAT-6 with a mutation disrupting the ATPase activity, can still promote cell death, suggesting that EAT-6 has a function in promoting cell death independent of its pumping activity. In addition, EAT-6 can promote cell death in the presence or absence of beta subunit NKB-1/NKB-2 in a cell-specific manner. And I also found KIN-2, a protein kinase A (cAMP-dependent protein kinase) regulatory subunit can promote programmed cell death in C. elegans. A typical mammalian protein kinase A is composed of two regulatory subunits and two catalytic subunits. C. elegans has one regulatory subunit, kin-2, and one catalytic subunit, kin-1. I found that loss of the regulatory subunit, kin-2 reduced cell death during embryogenesis and, in contrast, loss of the catalytic subunit kin-1 increased cell death during embryogenesis. In addition, overexpression of kin-2 promotes cell death. And I found that expression of eat-6 or kin-2 in dying cells rescued the cell death defeat in eat-6 or kin-2 mutants, indicating that eat-6 and kin-2 function in dying cells. And I found that eat-6 and kin-2 promote cell death through egl-1. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T13:20:59Z (GMT). No. of bitstreams: 1 ntu-102-R00b43005-1.pdf: 1348964 bytes, checksum: 79e3e87c6dcff7349fd96d2b5b957de2 (MD5) Previous issue date: 2013 | en |
dc.description.tableofcontents | 口試委員會審定書 ii
致謝 iii 中文摘要 iv ABSTRACT vi INTRODUCTION 4 MATERIALS AND METHODS 10 Strains and alleles 10 RNAi experiments 10 Plasmids construction 11 Transgenic and rescue experiments 12 RESULTS 14 Part1. Na+-K+ ATPase α subunit eat-6 14 The Na+-K+ ATPase α subunit eat-6 mutation reduces cell corpse numbers during embryogenesis 14 The Na+-K+ ATPase β subunit nkb-1, nkb-2 and nkb-3 mutations do not affect cell corpse numbers during embryogenesis 15 Excretory cell sister cell is missing in the eat-6(ad997) mutant 16 eat-6 RNAi or mutation enhances cell survival in the weak ced-3 mutants 16 eat-6 functions in dying cells 18 EAT-6 has the killing function and acts cell autonomously to kill cells 18 Pump dead EAT-6 still can promote cell death 19 NKB-1 or NKB-2 can partially suppress EAT-6 killing activity 20 EAT-6 is expressed in dying cells, pharynx, body-wall muscle, hypodermis, vulval muscle and touch neurons 21 eat-6 promotes cell death through the core cell death gene egl-1 and ced-4 22 Part2. Protein kinase A regulatory subunit kin-2 22 Protein kinase A regulatory subunit kin-2 mutation reduces cell corpse numbers during embryogenesis 22 Protein kinase A catalytic subunit kin-1 mutation enhances cell corpse numbers during embryogenesis 23 kin-2 RNAi enhances cell survival in weak ced-3 mutants 24 kin-2 functions in dying cells 25 KIN-2 has the killing function and acts cell autonomously to kill cells 25 KIN-2 is expressed in nerve ring 26 kin-2 promotes cell death through the core cell death gene egl-1 26 DISCUSSION 27 REFERENCES 32 TABLES and FIGURES 36 Table 1. eat-6 RNAi or mutation enhances cell survival in the weak ced-3 mutants 36 Table 2. Expression of eat-6 in dying cells rescues the cell corpse numbers in eat-6 mutant 37 Table 3. Overexpression of eat-6 promotes cell death 38 Table 4. Pump-dead EAT-6(R669Q) still can kill cells 39 Table 5. NKB-1 can partially suppress EAT-6 killing activity 40 Table 6. NKB-2 can partially suppress EAT-6 killing activity 41 Table 7. Overexpression of eat-6 can not kill cells in the absence of egl-1 42 Table 8. Overexpression of eat-6 can not kill cells in the absence of ced-4 43 Table 9. Overexpression of kin-2 rescues the cell corpse numbers in kin-2 mutant 44 Table 10. kin-2 RNAi enhances cell survival in the weak ced-3 mutants 45 Table 11. Expression of kin-2 in dying cells rescues the cell corpse numbers in kin-2 mutant 46 Table 12. Overexpression of kin-2 promotes cell death 47 Table 13. Overexpression of kin-2 can not kill cells in the absence of egl-1 48 Figure 1. The Na+-K+ ATPase α subunit eat-6 mutation reduces cell corpse numbers during embryogenesis 49 Figure 2. The Na+-K+ ATPase β subunit nkb-1, nkb-2 and nkb-3 mutations do not affect cell corpse numbers during embryogenesis 50 Figure 3. EAT-6 is expressed in pharynx, body-wall muscle, hypodermis, vulval muscle and touch neuron 51 Figure 4. eat-6 is expressed in cell corpse 52 Figure 5. kin-2 mutation reduces cell corpse numbers during embryogenesis 53 Figure 6. kin-1 mutation increases cell corpse numbers during embryogenesis 54 Figure 7. KIN-2 is expressed in nerve ring 55 Figure 8. Excretory cell sister cell is missing in the eat-6(ad997) mutant 56 Figure 9. Current model 57 | |
dc.language.iso | en | |
dc.title | 鈉鉀ATP酶和蛋白激酶A調控線蟲計劃性細胞死亡 | zh_TW |
dc.title | Na+-K+ ATPase and protein kinase A regulate programmed cell death in C. elegans | en |
dc.type | Thesis | |
dc.date.schoolyear | 101-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 潘俊良,陳俊宏 | |
dc.subject.keyword | 線蟲,計畫性細胞死亡,鈉鉀ATP?,蛋白激?A, | zh_TW |
dc.subject.keyword | C. elegans,programmed cell death,Na+-K+ ATPase,protein kinase A, | en |
dc.relation.page | 57 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2013-07-25 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 分子與細胞生物學研究所 | zh_TW |
顯示於系所單位: | 分子與細胞生物學研究所 |
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