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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 李財坤(Tsai-Kun Li) | |
| dc.contributor.author | An-Lin Peng | en |
| dc.contributor.author | 彭安林 | zh_TW |
| dc.date.accessioned | 2021-06-15T01:25:04Z | - |
| dc.date.available | 2014-09-15 | |
| dc.date.copyright | 2009-09-15 | |
| dc.date.issued | 2009 | |
| dc.date.submitted | 2009-07-23 | |
| dc.identifier.citation | Antony, S., Marchand, C., Stephen, A.G., Thibaut, L., Agama, K.K., Fisher, R.J., and Pommier, Y. (2007). Novel high-throughput electrochemiluminescent assay for identification of human tyrosyl-DNA phosphodiesterase (Tdp1) inhibitors and characterization of furamidine (NSC 305831) as an inhibitor of Tdp1. Nucleic Acids Res 35, 4474-4484.
Aplan, P.D. (2006). Chromosomal translocations involving the MLL gene: molecular mechanisms. DNA Repair (Amst) 5, 1265-1272. Austin, C.A., and Marsh, K.L. (1998). Eukaryotic DNA topoisomerase II beta. Bioessays 20, 215-226. Austin, C.A., Sng, J.H., Patel, S., and Fisher, L.M. (1993). Novel HeLa topoisomerase II is the II beta isoform: complete coding sequence and homology with other type II topoisomerases. Biochim Biophys Acta 1172, 283-291. Baguley, B.C., and Ferguson, L.R. (1998). Mutagenic properties of topoisomerase-targeted drugs. Biochim Biophys Acta 1400, 213-222. Baker, S.D., Wadkins, R.M., Stewart, C.F., Beck, W.T., and Danks, M.K. (1995). Cell cycle analysis of amount and distribution of nuclear DNA topoisomerase I as determined by fluorescence digital imaging microscopy. Cytometry 19, 134-145. Barthelmes, H.U., Habermeyer, M., Christensen, M.O., Mielke, C., Interthal, H., Pouliot, J.J., Boege, F., and Marko, D. (2004). TDP1 overexpression in human cells counteracts DNA damage mediated by topoisomerases I and II. J Biol Chem 279, 55618-55625. Burma, S., Chen, B.P., and Chen, D.J. (2006). Role of non-homologous end joining (NHEJ) in maintaining genomic integrity. DNA Repair (Amst) 5, 1042-1048. Caldecott, K.W., Tucker, J.D., Stanker, L.H., and Thompson, L.H. (1995). Characterization of the XRCC1-DNA ligase III complex in vitro and its absence from mutant hamster cells. Nucleic Acids Res 23, 4836-4843. Chaly, N., Chen, X., Dentry, J., and Brown, D.L. (1996). Organization of DNA topoisomerase II isotypes during the cell cycle of human lymphocytes and HeLa cells. Chromosome Res 4, 457-466. Champoux, J.J. (2001). DNA topoisomerases: structure, function, and mechanism. Annu Rev Biochem 70, 369-413. D'Arpa, P., Beardmore, C., and Liu, L.F. (1990). Involvement of nucleic acid synthesis in cell killing mechanisms of topoisomerase poisons. Cancer Res 50, 6919-6924. D'Arpa, P., Machlin, P.S., Ratrie, H., 3rd, Rothfield, N.F., Cleveland, D.W., and Earnshaw, W.C. (1988). cDNA cloning of human DNA topoisomerase I: catalytic activity of a 67.7-kDa carboxyl-terminal fragment. Proc Natl Acad Sci U S A 85, 2543-2547. Daroui, P., Desai, S.D., Li, T.K., Liu, A.A., and Liu, L.F. (2004). Hydrogen peroxide induces topoisomerase I-mediated DNA damage and cell death. J Biol Chem 279, 14587-14594. Davies, D.R., Interthal, H., Champoux, J.J., and Hol, W.G. (2002a). The crystal structure of human tyrosyl-DNA phosphodiesterase, Tdp1. Structure 10, 237-248. Davies, D.R., Interthal, H., Champoux, J.J., and Hol, W.G. (2002b). Insights into substrate binding and catalytic mechanism of human tyrosyl-DNA phosphodiesterase (Tdp1) from vanadate and tungstate-inhibited structures. J Mol Biol 324, 917-932. Davies, D.R., Interthal, H., Champoux, J.J., and Hol, W.G. (2003). Crystal structure of a transition state mimic for Tdp1 assembled from vanadate, DNA, and a topoisomerase I-derived peptide. Chem Biol 10, 139-147. de Murcia, G., and Menissier de Murcia, J. (1994). Poly(ADP-ribose) polymerase: a molecular nick-sensor. Trends Biochem Sci 19, 172-176. Debethune, L., Kohlhagen, G., Grandas, A., and Pommier, Y. (2002). Processing of nucleopeptides mimicking the topoisomerase I-DNA covalent complex by tyrosyl-DNA phosphodiesterase. Nucleic Acids Res 30, 1198-1204. Desai, S.D., Li, T.K., Rodriguez-Bauman, A., Rubin, E.H., and Liu, L.F. (2001). Ubiquitin/26S proteasome-mediated degradation of topoisomerase I as a resistance mechanism to camptothecin in tumor cells. Cancer Res 61, 5926-5932. Desai, S.D., Liu, L.F., Vazquez-Abad, D., and D'Arpa, P. (1997). Ubiquitin-dependent destruction of topoisomerase I is stimulated by the antitumor drug camptothecin. J Biol Chem 272, 24159-24164. Desai, S.D., Zhang, H., Rodriguez-Bauman, A., Yang, J.M., Wu, X., Gounder, M.K., Rubin, E.H., and Liu, L.F. (2003). Transcription-dependent degradation of topoisomerase I-DNA covalent complexes. Mol Cell Biol 23, 2341-2350. El-Khamisy, S.F., Saifi, G.M., Weinfeld, M., Johansson, F., Helleday, T., Lupski, J.R., and Caldecott, K.W. (2005). Defective DNA single-strand break repair in spinocerebellar ataxia with axonal neuropathy-1. Nature 434, 108-113. Fan, J.R., Peng, A.L., Chen, H.C., Lo, S.C., Huang, T.H., and Li, T.K. (2008). Cellular processing pathways contribute to the activation of etoposide-induced DNA damage responses. DNA Repair (Amst) 7, 452-463. Felix, C.A. (1998). Secondary leukemias induced by topoisomerase-targeted drugs. Biochim Biophys Acta 1400, 233-255. Felix, C.A., Kolaris, C.P., and Osheroff, N. (2006). Topoisomerase II and the etiology of chromosomal translocations. DNA Repair (Amst) 5, 1093-1108. Gangloff, S., Lieber, M.R., and Rothstein, R. (1994). Transcription, topoisomerases and recombination. Experientia 50, 261-269. Gellert, M., Mizuuchi, K., O'Dea, M.H., and Nash, H.A. (1976). DNA gyrase: an enzyme that introduces superhelical turns into DNA. Proc Natl Acad Sci U S A 73, 3872-3876. Hanai, R., Caron, P.R., and Wang, J.C. (1996). Human TOP3: a single-copy gene encoding DNA topoisomerase III. Proc Natl Acad Sci U S A 93, 3653-3657. Hiasa, H., Yousef, D.O., and Marians, K.J. (1996). DNA strand cleavage is required for replication fork arrest by a frozen topoisomerase-quinolone-DNA ternary complex. J Biol Chem 271, 26424-26429. Hirano, R., Interthal, H., Huang, C., Nakamura, T., Deguchi, K., Choi, K., Bhattacharjee, M.B., Arimura, K., Umehara, F., Izumo, S., et al. (2007). Spinocerebellar ataxia with axonal neuropathy: consequence of a Tdp1 recessive neomorphic mutation? EMBO J 26, 4732-4743. Howard, M.T., Neece, S.H., Matson, S.W., and Kreuzer, K.N. (1994). Disruption of a topoisomerase-DNA cleavage complex by a DNA helicase. Proc Natl Acad Sci U S A 91, 12031-12035. Hsiang, Y.H., Lihou, M.G., and Liu, L.F. (1989). Arrest of replication forks by drug-stabilized topoisomerase I-DNA cleavable complexes as a mechanism of cell killing by camptothecin. Cancer Res 49, 5077-5082. Huang, T.T., Wuerzberger-Davis, S.M., Seufzer, B.J., Shumway, S.D., Kurama, T., Boothman, D.A., and Miyamoto, S. (2000). NF-kappaB activation by camptothecin. A linkage between nuclear DNA damage and cytoplasmic signaling events. J Biol Chem 275, 9501-9509. Interthal, H., Chen, H.J., Kehl-Fie, T.E., Zotzmann, J., Leppard, J.B., and Champoux, J.J. (2005). SCAN1 mutant Tdp1 accumulates the enzyme--DNA intermediate and causes camptothecin hypersensitivity. EMBO J 24, 2224-2233. Interthal, H., Pouliot, J.J., and Champoux, J.J. (2001). The tyrosyl-DNA phosphodiesterase Tdp1 is a member of the phospholipase D superfamily. Proc Natl Acad Sci U S A 98, 12009-12014. Jenkins, J.R., Ayton, P., Jones, T., Davies, S.L., Simmons, D.L., Harris, A.L., Sheer, D., and Hickson, I.D. (1992). Isolation of cDNA clones encoding the beta isozyme of human DNA topoisomerase II and localisation of the gene to chromosome 3p24. Nucleic Acids Res 20, 5587-5592. Katyal, S., el-Khamisy, S.F., Russell, H.R., Li, Y., Ju, L., Caldecott, K.W., and McKinnon, P.J. (2007). TDP1 facilitates chromosomal single-strand break repair in neurons and is neuroprotective in vivo. EMBO J 26, 4720-4731. Leiros, I., Secundo, F., Zambonelli, C., Servi, S., and Hough, E. (2000). The first crystal structure of a phospholipase D. Structure 8, 655-667. Leppard, J.B., and Champoux, J.J. (2005). Human DNA topoisomerase I: relaxation, roles, and damage control. Chromosoma 114, 75-85. Li, T.K., and Liu, L.F. (2001). Tumor cell death induced by topoisomerase-targeting drugs. Annu Rev Pharmacol Toxicol 41, 53-77. Liao, Z., Thibaut, L., Jobson, A., and Pommier, Y. (2006). Inhibition of human tyrosyl-DNA phosphodiesterase by aminoglycoside antibiotics and ribosome inhibitors. Mol Pharmacol 70, 366-372. Lieber, M.R., Ma, Y., Pannicke, U., and Schwarz, K. (2003). Mechanism and regulation of human non-homologous DNA end-joining. Nat Rev Mol Cell Biol 4, 712-720. Lin, C.P., Ban, Y., Lyu, Y.L., Desai, S.D., and Liu, L.F. (2008). A ubiquitin-proteasome pathway for the repair of topoisomerase I-DNA covalent complexes. J Biol Chem 283, 21074-21083. Mao, Y., Desai, S.D., Ting, C.Y., Hwang, J., and Liu, L.F. (2001). 26 S proteasome-mediated degradation of topoisomerase II cleavable complexes. J Biol Chem 276, 40652-40658. Mistry, A.R., Felix, C.A., Whitmarsh, R.J., Mason, A., Reiter, A., Cassinat, B., Parry, A., Walz, C., Wiemels, J.L., Segal, M.R., et al. (2005). DNA topoisomerase II in therapy-related acute promyelocytic leukemia. N Engl J Med 352, 1529-1538. Morham, S.G., Kluckman, K.D., Voulomanos, N., and Smithies, O. (1996). Targeted disruption of the mouse topoisomerase I gene by camptothecin selection. Mol Cell Biol 16, 6804-6809. Ng, S.W., Liu, Y., Hasselblatt, K.T., Mok, S.C., and Berkowitz, R.S. (1999). A new human topoisomerase III that interacts with SGS1 protein. Nucleic Acids Res 27, 993-1000. Nitiss, K.C., Malik, M., He, X., White, S.W., and Nitiss, J.L. (2006). Tyrosyl-DNA phosphodiesterase (Tdp1) participates in the repair of Top2-mediated DNA damage. Proc Natl Acad Sci U S A 103, 8953-8958. Plo, I., Liao, Z.Y., Barcelo, J.M., Kohlhagen, G., Caldecott, K.W., Weinfeld, M., and Pommier, Y. (2003). Association of XRCC1 and tyrosyl DNA phosphodiesterase (Tdp1) for the repair of topoisomerase I-mediated DNA lesions. DNA Repair (Amst) 2, 1087-1100. Pommier, Y., Barcelo, J.M., Rao, V.A., Sordet, O., Jobson, A.G., Thibaut, L., Miao, Z.H., Seiler, J.A., Zhang, H., Marchand, C., et al. (2006). Repair of topoisomerase I-mediated DNA damage. Prog Nucleic Acid Res Mol Biol 81, 179-229. Pouliot, J.J., Yao, K.C., Robertson, C.A., and Nash, H.A. (1999). Yeast gene for a Tyr-DNA phosphodiesterase that repairs topoisomerase I complexes. Science 286, 552-555. Raymond, A.C., Rideout, M.C., Staker, B., Hjerrild, K., and Burgin, A.B., Jr. (2004). Analysis of human tyrosyl-DNA phosphodiesterase I catalytic residues. J Mol Biol 338, 895-906. Raymond, A.C., Staker, B.L., and Burgin, A.B., Jr. (2005). Substrate specificity of tyrosyl-DNA phosphodiesterase I (Tdp1). J Biol Chem 280, 22029-22035. Shao, R.G., Cao, C.X., Zhang, H., Kohn, K.W., Wold, M.S., and Pommier, Y. (1999). Replication-mediated DNA damage by camptothecin induces phosphorylation of RPA by DNA-dependent protein kinase and dissociates RPA:DNA-PK complexes. EMBO J 18, 1397-1406. Shea, M.E., and Hiasa, H. (2000). Distinct effects of the UvrD helicase on topoisomerase-quinolone-DNA ternary complexes. J Biol Chem 275, 14649-14658. Stuckey, J.A., and Dixon, J.E. (1999). Crystal structure of a phospholipase D family member. Nat Struct Biol 6, 278-284. Subramanian, D., Kraut, E., Staubus, A., Young, D.C., and Muller, M.T. (1995). Analysis of topoisomerase I/DNA complexes in patients administered topotecan. Cancer Res 55, 2097-2103. Taagepera, S., Rao, P.N., Drake, F.H., and Gorbsky, G.J. (1993). DNA topoisomerase II alpha is the major chromosome protein recognized by the mitotic phosphoprotein antibody MPM-2. Proc Natl Acad Sci U S A 90, 8407-8411. Takashima, H., Boerkoel, C.F., John, J., Saifi, G.M., Salih, M.A., Armstrong, D., Mao, Y., Quiocho, F.A., Roa, B.B., Nakagawa, M., et al. (2002). Mutation of TDP1, encoding a topoisomerase I-dependent DNA damage repair enzyme, in spinocerebellar ataxia with axonal neuropathy. Nat Genet 32, 267-272. Taylor, R.M., Wickstead, B., Cronin, S., and Caldecott, K.W. (1998). Role of a BRCT domain in the interaction of DNA ligase III-alpha with the DNA repair protein XRCC1. Curr Biol 8, 877-880. Tomicic, M.T., Christmann, M., and Kaina, B. (2005). Topotecan-triggered degradation of topoisomerase I is p53-dependent and impacts cell survival. Cancer Res 65, 8920-8926. Tsao, Y.P., Russo, A., Nyamuswa, G., Silber, R., and Liu, L.F. (1993). Interaction between replication forks and topoisomerase I-DNA cleavable complexes: studies in a cell-free SV40 DNA replication system. Cancer Res 53, 5908-5914. Wan, S., Capasso, H., and Walworth, N.C. (1999). The topoisomerase I poison camptothecin generates a Chk1-dependent DNA damage checkpoint signal in fission yeast. Yeast 15, 821-828. Wang, J.C. (1971). Interaction between DNA and an Escherichia coli protein omega. J Mol Biol 55, 523-533. Wang, J.C. (1985). DNA topoisomerases: nature's solution to the topological ramifications of the double-helix structure of DNA. Harvey Lect 81, 93-110. Wang, J.C. (2002). Cellular roles of DNA topoisomerases: a molecular perspective. Nat Rev Mol Cell Biol 3, 430-440. Warburton, P.E., and Earnshaw, W.C. (1997). Untangling the role of DNA topoisomerase II in mitotic chromosome structure and function. Bioessays 19, 97-99. Whitehouse, C.J., Taylor, R.M., Thistlethwaite, A., Zhang, H., Karimi-Busheri, F., Lasko, D.D., Weinfeld, M., and Caldecott, K.W. (2001). XRCC1 stimulates human polynucleotide kinase activity at damaged DNA termini and accelerates DNA single-strand break repair. Cell 104, 107-117. Wu, J., and Liu, L.F. (1997). Processing of topoisomerase I cleavable complexes into DNA damage by transcription. Nucleic Acids Res 25, 4181-4186. Wyman, C., and Kanaar, R. (2006). DNA double-strand break repair: all's well that ends well. Annu Rev Genet 40, 363-383. Xiao, H., Mao, Y., Desai, S.D., Zhou, N., Ting, C.Y., Hwang, J., and Liu, L.F. (2003). The topoisomerase IIbeta circular clamp arrests transcription and signals a 26S proteasome pathway. Proc Natl Acad Sci U S A 100, 3239-3244. Yang, S.W., Burgin, A.B., Jr., Huizenga, B.N., Robertson, C.A., Yao, K.C., and Nash, H.A. (1996). A eukaryotic enzyme that can disjoin dead-end covalent complexes between DNA and type I topoisomerases. Proc Natl Acad Sci U S A 93, 11534-11539. Yang, X., Li, W., Prescott, E.D., Burden, S.J., and Wang, J.C. (2000). DNA topoisomerase IIbeta and neural development. Science 287, 131-134. Zhang, A., Lyu, Y.L., Lin, C.P., Zhou, N., Azarova, A.M., Wood, L.M., and Liu, L.F. (2006). A protease pathway for the repair of topoisomerase II-DNA covalent complexes. J Biol Chem 281, 35997-36003. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/42829 | - |
| dc.description.abstract | 引發拓樸異構酶可切性複合體(TOPcc)被認為是有效的抗微生物以及抗癌策略，因其可以造成有蛋白質連結在3端或5端DNA斷點上的特殊DNA傷害。人類酪胺酸DNA磷脂酶(human tyrosyl-DNA phosphodiesterase 1, hTDP1)藉由其水解3’磷酸二脂鍵的能力而參與在處理以及修復TOP1cc所造成的DNA傷害過程中。然而，hTDP1在處理TOP2cc中是否扮演腳色，以及和TOPcc的其他細胞處理途徑(複製起始的處理：RIP，和轉錄起始的處理：TIP)之間的關係，仍不清楚。因此，本論文利用藥物調控、RNA干擾技術，和強制蛋白質表現等策略來探討hTDP1對TOP2cc引發的DNA傷害及反應之影響。
我們分別在HCT116細胞株和BT-474細胞中降低hTDP1的蛋白質表現；和控制組細胞比較，發現hTDP1缺乏的細胞對於拓樸異構酶II藥物所造成的DNA斷點較多，且對於藥物的敏感性也顯著增加；相反地，在293細胞以及hTDP1缺乏的細胞中強制其蛋白質表現則可有效抵制上述表徵。此外，clonogenic assay的結果也支持著hTDP1對於TOP2cc引發之細胞死亡有著保護性的腳色。而hTDP1的缺乏顯著地增加了細胞對於TOP1和TOP2藥物引起之NHEJ事件，更加支持hTDP1可能在細胞處理TOP2cc的過程中佔有一席之地。 接下來，為了探討hTDP1和RIP/TIP的關係，我們研究兩者對TOP1cc的差異。首先，我們發現缺乏hTDP1並不影響TOP1cc的降解，表示hTDP1可能作用於蛋白體降解(proteasome degradation)之後，或是作用於和TIP路徑平行的RIP路徑。根據實驗室先前對於不同處理途徑所引發特定的TOP1藥物造成之DNA傷害反應，相較於控制組細胞，我們發現hTDP1缺乏的細胞處理TOP1藥物後能引發較高程度的Chk2和p53活化(TIP引發的傷害訊息分子)，而RIP引發的傷害訊息分子，DNA-PK和RPA的活化則沒有顯著差異：使用藥物分別抑制複製和轉錄也得到一致的結論。因此，hTDP1可能經由TIP路徑活化TOP1藥物所引發下游的DNA傷害反應。總括而論，本研究可提供合併療法在機制上的解釋，並對於以TOP作為抗癌標的的治療方針的效益提升貢獻了可能的幫助。 | zh_TW |
| dc.description.abstract | Formation of topoisomerase cleavable complex (TOPcc), in an unique form of protein-linked 3’- or 5’-DNA breakage, represents as an effective antibiotic and anti-cancer strategy. Human tyrosyl-DNA phosphodiesterase1 (hTDP1) has been implied in TOP1cc processing and repair through the activity of hydrolyzing 3’-phosphotyrosyl bond between TOP1 and DNA. However, the role of hTDP1 in TOP2cc processing and the relationship among hTDP1, TIP and RIP pathways remain unclear. Toward this aim, a combination of pharmacological modulation, RNAi technology and forced expression approach was utilized to investigate the impact of hTDP1 on TOP2cc-mediated DNA breakage and damage responses (DDR).
Two pairs of hTDP1-knockdown and control cells had been successfully established in HCT116 cells and BT-474 cells. Our results showed hTDP1-deficient cells exhibited a higher extent of DNA breakage and hypersensitivity to TOP2-targeting drugs. While forced expression of hTDP1 in 293 cells and hTDP1-deficient cells could reduce TOP2-mediated DNA damage and counteract these hTDP1-deficient phenotypes. The clonogenic assay additionally supported the protective role of hTDP1 in TOP2cc-mediated cell killing. Moreover, the increased rate of CPT- and VP16-induced NHEJ-mediated plasmid integration in hTDP1-deficient cells also consistent with the potential involvement of hTDP1 in the processing of TOP2cc. In regarding to the relationship between hTDP1 and cellular processing pathways of TOP1cc, we first observed that CPT-induced downregulation of TOP1cc was not affected in hTDP1-deficient cells. Two possible explanations were offered: (i) hTDP1 could be involved in the downstream of proteasome degradation. (ii) hTDP1 could be involved in RIP, which is parallel with proteasome degradation. Based on our previous study on distinct activation of CPT-induced DNA damage responses (DDR) via two processing pathways, we further observed that phosphorylations of Chk2 and p53 (TIP-related signals) rather than DNA-PK, RPA (RIP-related signals) were increased upon CPT treatment in hTDP1-deficient cells. Similar conclusion was also obtained from pharmacological modulation studies. Therefore, our data suggest a potential contribution of hTDP1 to CPT-induced DDR through TIP pathway. In sum, our studies might therefore provide mechanistic explanations for these combination effects and further helps for the development of TOP-based therapeutic intervention. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-15T01:25:04Z (GMT). No. of bitstreams: 1 ntu-98-R96445101-1.pdf: 2089546 bytes, checksum: d1a5780182349d7f63c59b3f0d6bc39f (MD5) Previous issue date: 2009 | en |
| dc.description.tableofcontents | 口試委員會審定書 ……………………………………………………i
中文摘要 ………………………………………………………………ii ABSTRACT………………………………………………………………iii CONTENTS…………………………………………………………………v INTRODUCTION……………………………………………………………1 1. DNA topoisomerase ……………………………………………1 1.1 Type I DNA topoisomerase …………………………………2 1.2 Type II DNA topoisomerase ……………………………… 2 1.3 Cellular processing pathways of topoisomerase-mediated DNA damage…………………………………………………4 1.3.1 Replication-initiated processing (RIP) pathway …………………………………………………………………………5 1.3.2 Transcription-initiated processing (TIP) pathway ……………………………………………………………………………5 2. Tyrosyl-DNA phosphodiesterase 1 …………………………6 2.1 Structure ……………………………………………………7 2.2 Catalytic mechanism ………………………………………8 2.3 Substrate binding and accessible substrate …………8 2.4 Spinocerebellar ataxia with axonal neuropathy (SCAN1)……………………………………………………………………………9 2.5 TDP1-dependent DNA repair pathway ……………………10 SPECIFIC AIMS…………………………………………………………12 MATERIALS and METHODS ………………………………………………14 - Chemicals and cell culture ……………………………… 14 - RNA interference(RNAi) knockdown technology …………14 - Plasmids and cell transfection …………………………14 - Antibodies and immunoblotting analysis ………………15 - Quantitative measurement and statistic analysis ……16 - In vivo complex enzyme (ICE) assay ……………………16 - Alkaline single cell gel electroforesis (Comet) assay …………………………………………………………………………17 - Cytotoxicity assay – MTT analysis and colony formation assay ……………………………………………………18 - Plasmid integration assay …………………………………18 - Immunofluorescence assay (IFA)……………………………19 RESULTS…………………………………………………………………20 DISCUSSION………………………………………………………………29 REFERENCES………………………………………………………………35 FIGURES…………………………………………………………………40 APPENDIX………………………………………………………………65 | |
| dc.language.iso | en | |
| dc.subject | DNA磷脂酶 | zh_TW |
| dc.subject | 人類酪胺酸 | zh_TW |
| dc.subject | 可切性複合體 | zh_TW |
| dc.subject | 拓樸異構酶 | zh_TW |
| dc.subject | cleavable complex | en |
| dc.subject | hTDP1 | en |
| dc.subject | topoisomerase | en |
| dc.title | 探討人類酪胺酸DNA磷脂酶在處理拓樸異構酶可切性複合體上所扮演的角色 | zh_TW |
| dc.title | Study on Roles of Human Tyrosyl-DNA Phosphodiesterase 1 in processing of Topoisomerase Cleavable Complexes | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 97-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 鄧述諄(Shu-Chun Teng),詹迺立(Nei-Li Chan) | |
| dc.subject.keyword | 人類酪胺酸,DNA磷脂酶,拓樸異構酶,可切性複合體, | zh_TW |
| dc.subject.keyword | hTDP1,topoisomerase,cleavable complex, | en |
| dc.relation.page | 66 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2009-07-23 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 微生物學研究所 | zh_TW |
| 顯示於系所單位： | 微生物學科所 | |
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