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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 吳益群(Yi-Chun Wu) | |
dc.contributor.author | Chan-Hsien Yeh | en |
dc.contributor.author | 葉蟬嫻 | zh_TW |
dc.date.accessioned | 2021-06-15T04:45:37Z | - |
dc.date.available | 2015-08-11 | |
dc.date.copyright | 2010-08-11 | |
dc.date.issued | 2010 | |
dc.date.submitted | 2010-08-05 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/45755 | - |
dc.description.abstract | 在細胞進行有絲分裂過程中,染色體的濃縮與分離和細胞質分裂若是發生異常,就會破壞基因體的完整性並造成生物體的死亡或是癌化。線蟲裡的condensin和AIR-2蛋白,對於細胞分裂能夠正常執行是必需的;TLK-1已被證實是AIR-2的受質及活化物,在AIR-2與另一個活化物ICP-1結合的情況下,能參與它們的作用共同確保染色質分裂能正常進行。然而TLK-1是否有參與染色質濃縮或是細胞質分裂仍是未知問題。在論文裡,我們使用顯微鏡追蹤分析tlk-1突變株發現,TLK-1不只作用在染色體分離過程,也作用在染色體濃縮和細胞質分裂。從結果顯示,TLK-1以不同於ICP-1的作用方式參與染色體濃縮和細胞質分裂,既不影響AIR-2和condensin與染色體結合的能力,整個kinetochore的結構和功能也是正常的,只有染色體的排列有問題。更甚者,TLK-1在細胞分裂過程中的位置分佈也與AIR-2和ICP-1不同,在核膜破裂後TLK-1散布到整個細胞中,而不是結合到染色體上,這也與之前TLK-1的研究結果不一樣;由此可知,TLK-1雖是AIR-2的活化蛋白,但它是以不同於ICP-1的方式和AIR-2共同作用。而TLK-1影響細胞質分裂的分式,則是透過調控AIR-2在細胞分裂末期的分佈,使之能坐落到midzone microtubule以促進細胞質分裂完成。除此之外,TLK-1還可能透過磷酸化HCP-3的模式,使染色體在進行濃縮時能形成正確的結構。最後透過yeast two-hybrid screen,我們發現了一個新的可與TLK-1共同作用的蛋白,DLC-1。TLK-1可能與DLC-1共同影響microtubule和kinetochore的交互作用。 | zh_TW |
dc.description.abstract | Defects in chromosome condensation, segregation or cytokinesis during mitosis disrupt genome integrity and cause organismal death or tumorigenesis. The conserved kinase protein AIR-2/Aurora B is required for normal execution of all these important mitotic events in C. elegans. TLK-1 has been recently shown to be a substrate and activator of AIR-2 in the presence of another AIR-2 activator ICP-1/INCENP, and to cooperate with AIR-2 to ensure proper mitotic chromosome segregation. However, whether TLK-1 may contribute to chromosome condensation or cytokinesis is unclear. A time-lapse microscopy analysis showed that tlk-1 mutants are defective in chromosome condensation and cytokinesis, in addition to chromosome segregation, during mitosis. Our data indicate that TLK-1 contributes to chromosome condensation and segregation, at least in part, in a manner that is distinct from the ICP-1-mediated mechanism and does not involve loading AIR-2 and condensin complexes to mitotic chromosomes. The overall structure of kinetochore is also normal accompanies with the chromosome alignment defect during metaphase in tlk-1 mutant. Moreover, the localization pattern of TLK-1 is different from the previous data and AIR-2/ICP-1 complex, which TLK-1 diffused to cytoplasm after nuclear envelope broken down. This reveals the differences in dynamic regulation and association of TLK-1 and ICP-1 towards AIR-2 in vivo. Finally, TLK-1 functions in cytokinesis by localizing AIR-2 to the midzone microtubules. Otherwise, TLK-1 may have function in chromosome architecture during chromosome condensation via phosphorylating HCP-3. And via yeast two-hybrid screen, we found a new TLK-1 interacting protein, DLC-1, which may cooperate to regulate the microtubule and kinetochore interaction during mitosis. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T04:45:37Z (GMT). No. of bitstreams: 1 ntu-99-D92B43004-1.pdf: 4055317 bytes, checksum: 78ca6d637b935c8f3d2b15c80befa9e1 (MD5) Previous issue date: 2010 | en |
dc.description.tableofcontents | 1.Introduction …… 1
1.1 Condensin and Topoisomerase II are two important chromosome scaffold proteins …… 1 1.2 Topoisomerase II functions in chromatid resolution …… 2 1.3 Two condensin complexes:condensin I and II …… 3 1.4 Centromeric condensin complexes …… 5 1.5 Regulation of condensin complexes …… 5 1.6 The really role of condensin complexes during chromosome condensation …… 7 1.7 The chromosome passenger complex …… 9 1.8 The functions of Aurora B kinase at centromere …… 10 1.9 The functions of Aurora B kinase at midzone microtubules…… 11 1.10 The function of Aurora B kinase at anaphase bridged chromosome …… 13 1.11 The overall functions of TLK during interphase …… 13 1.12 The overall functions of TLK during mitosis…… 17 1.13 The TLK functions in unicellular protozoan …… 18 1.14 What we want to know about TLK-1 in C. elegans …… 19 1.15 What we have known in our experiments …… 20 2.Materials and Methods …… 22 2.1 Strains and alleles …… 22 2.2 RNAi experiments …… 22 2.3 Live imaging …… 23 2.4 ostaining …… 23 2.5 Yeast two-hybrid assay …… 25 2.6 Kinase assay …… 25 2.7 DAPI staining …… 26 2.8 Construct of tlk-1(kd) gene …… 26 3.Results …… 27 3.1 The tlk-1(RNAi) embryos are defective in chromosome architecture, segregation and alignment …… 27 3.2 The chromosome segregation defect was observed prior to the chromosome condensation defect in tlk-1(RNAi) embryo 28 3.3 tlk-1 is important for chromosome condensation and cytokinesis …… 28 3.4 TLK-1 is constitutively present throughout cell cycle30 3.5 AIR-2 fails to localize to midzone microtubules at anaphase and telophase in the tlk-1(RNAi) mutant …… 31 3.6 TLK-1 may not mediate chromosome segregation by affecting CAPG-1 and CAPG-2 localization to chromosomes 32 3.7 The overall structure of kinetochore appears normal in tlk-1(RNAi) embryos …… 35 3.8 HCP-3 is the substrate of TLK-1 and may be involved in chromosome condensation during mitosis …… 36 3.9 DLC-1 could interact with TLK-1 independently of TLK-1 kinase activity …… 37 3.10 A deletion in tlk-1 results in a mitotic defect in post-embryonic development …… 38 4.Discussion …… 40 4.1 TLK-1 functions in chromosome condensation, segregation and cytokinesis may be independent with each other …… 40 4.2 TLK-1 and ICP-1/INCENP are two distinct substrate activator of AIR-2/Aurora B kinase …… 41 4.3 TLK-1 may have function in cytokinesis by regulating the localization of AIR-2 at spindle midzone during anaphase and telophase …… 41 4.4 TLK-1 may have function in chromosome architecture during chromosome condensation via phosphorylating HCP-3 42 4.5 TLK-1 may function with dynein in lateral binding of microtubules to the kinetochore …… 43 5.References …… 45 6.List of Figures and Table …… 63 Figure 1. Chromosome segregation and condensation defects in tlk-1(RNAi) embryo …… 63 Figure 2. Diagram of fixed cell lineage during early embryonic development in C. elegans …… 64 Figure 3. The time-course analysis of the tlk-1 chromosome segregation defect …… 65 Figure 4. The chromosome segregation defect in tlk-1(RNAi) embryos between 15- to 28-cell stage …… 66 Figure 5. Chromosome condensation appears normal until 28-cell stage in tlk-1 mutant …… 67 Figure 6. The time-course analysis of the tlk-1 chromosome condensation defect …… 68 Figure 7. The time-course analysis of the tlk-1 cytokinesis defect …… 69 Figure 8. The cell cycle-specific localization pattern of TLK-1 …… 70 Figure 9. No AIR-2 localization to midzone microtubules was detected in tlk-1 cytokinesis-defective cells in anaphase and telophase …… 71 Figure 10. CAPG-1 localization in wild-type and tlk-1 embryos …… 72 Figure 11. CAPG-2 localization in wild-type and tlk-1 embryos …… 73 Figure 12. KNL-2 localization in wild-type and tlk-1 embryos …… 74 Figure 13. MIS-12 localization in wild-type and tlk-1 embryos …… 75 Figure 14. KBP-5 localization in wild-type and tlk-1 embryos …… 76 Figure 15. HCP-3 is a substrate of TLK-1 in an in vitro kinase assay …… 77 Figure 16. DLC-1 interacts with TLK-1 but is not its in vitro substrate …… 78 Figure 17. A deletion mutation in tlk-1 results in both mitosis and meiosis defects …… 79 Table 1. Distance between spindle poles at metaphase in embryonic cells …… 80 | |
dc.language.iso | en | |
dc.title | 分析線蟲的TLK-1蛋白在細胞分裂中對DNA濃縮與分離和在細胞質分裂中的功能 | zh_TW |
dc.title | TLK-1 functions in DNA condensation, segregation and cytokinesis in Caenorhaditis elegans | en |
dc.type | Thesis | |
dc.date.schoolyear | 98-2 | |
dc.description.degree | 博士 | |
dc.contributor.oralexamcommittee | 黃偉邦,董桂書,李士傑,廖秀娟 | |
dc.subject.keyword | 有絲分裂,細胞質分裂,染色體濃縮,染色體分離,線蟲, | zh_TW |
dc.subject.keyword | tlk-1,cytokinesis,condensation,AIR-2,Aurora B,Caenorhabditis elegans, | en |
dc.relation.page | 80 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2010-08-06 | |
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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