請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/26523
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 陳宏文 | |
dc.contributor.author | Meng-Hsiu Chiang | en |
dc.contributor.author | 江孟修 | zh_TW |
dc.date.accessioned | 2021-06-08T07:13:38Z | - |
dc.date.copyright | 2008-08-06 | |
dc.date.issued | 2008 | |
dc.date.submitted | 2008-07-29 | |
dc.identifier.citation | Akiyama Y, Hosoya T, Poole AM, Hotta Y. (1996) The gcm-motif: a novel DNA-binding motif conserved in Drosophila and mammals. Proc Natl Acad Sci U S A., 93, 14912-14916.
Alfonso TB, Jones BW. (2002) gcm2 promotes glial cell differentiation and is required with glial cells missing for macrophage development in Drosophila. Dev Biol., 248, 369-383. Anderson DJ, Stone J, Lum R, Linial ML. (1995) The packaging phenotype of the SE21Q1b provirus is related to high proviral expression and not trans-acting factors. J Virol., 69, 7319-7323. Anson-Cartwright L, Dawson K, Holmyard D, Fisher SJ, Lazzarini RA, Cross JC. (2000) The glial cells missing-1 protein is essential for branching morphogenesis in the chorioallantoic placenta. Nat Genet., 25, 311-314. Basyuk E, Cross JC, Corbin J, Nakayama H, Hunter P, Nait-Oumesmar B, Lazzarini RA. (1999) Murine Gcm1 gene is expressed in a subset of placental trophoblast cells. Dev Dyn., 214, 303-311. Bailly V, Lamb J, Sung P, Prakash S, Prakash L. (1994) Specific complex formation between yeast RAD6 and RAD18 proteins: a potential mechanism for targeting RAD6 ubiquitin-conjugating activity to DNA damage sites. Genes Dev., 8, 811-820. Bernardoni R, Kammerer M, Vonesch JL, Giangrande A. (1999) Gliogenesis depends on glide/gcm through asymmetric division of neuroglioblasts. Dev Biol., 216, 265-275. Calestani C, Rast JP, Davidson EH. (2003) Isolation of pigment cell specific genes in the sea urchin embryo by differential macroarray screening. Development., 130, 4587-4596. Cary PD, King DS, Crane-Robinson C, Bradbury EM, Rabbani A, Goodwin GH, Johns EW. (1980) Structural studies on two high-mobility-group proteins from calf thymus, HMG-14 and HMG-20 (ubiquitin), and their interaction with DNA. Eur J Biochem., 112, 577-580. Castellucci M, Kaufmann P, Bischof P. (1990) Extracellular matrix influences hormone and protein production by human chorionic villi. Cell Tissue Res., 262, 135-142. Chang CW, Chuang HC, Yu C, Yao TP, Chen H. (2005) Stimulation of GCMa transcriptional activity by cyclic Akiyama Y, Hosoya T, Poole AM, Hotta Y. (1996) The gcm-motif: a novel DNA-binding motif conserved in Drosophila and mammals. Proc Natl Acad Sci U S A., 93, 14912-14916. Alfonso TB, Jones BW. (2002) gcm2 promotes glial cell differentiation and is required with glial cells missing for macrophage development in Drosophila. Dev Biol., 248, 369-383. Anderson DJ, Stone J, Lum R, Linial ML. (1995) The packaging phenotype of the SE21Q1b provirus is related to high proviral expression and not trans-acting factors. J Virol., 69, 7319-7323. Anson-Cartwright L, Dawson K, Holmyard D, Fisher SJ, Lazzarini RA, Cross JC. (2000) The glial cells missing-1 protein is essential for branching morphogenesis in the chorioallantoic placenta. Nat Genet., 25, 311-314. Basyuk E, Cross JC, Corbin J, Nakayama H, Hunter P, Nait-Oumesmar B, Lazzarini RA. (1999) Murine Gcm1 gene is expressed in a subset of placental trophoblast cells. Dev Dyn., 214, 303-311. Bailly V, Lamb J, Sung P, Prakash S, Prakash L. (1994) Specific complex formation between yeast RAD6 and RAD18 proteins: a potential mechanism for targeting RAD6 ubiquitin-conjugating activity to DNA damage sites. Genes Dev., 8, 811-820. Bernardoni R, Kammerer M, Vonesch JL, Giangrande A. (1999) Gliogenesis depends on glide/gcm through asymmetric division of neuroglioblasts. Dev Biol., 216, 265-275. Calestani C, Rast JP, Davidson EH. (2003) Isolation of pigment cell specific genes in the sea urchin embryo by differential macroarray screening. Development., 130, 4587-4596. Cary PD, King DS, Crane-Robinson C, Bradbury EM, Rabbani A, Goodwin GH, Johns EW. (1980) Structural studies on two high-mobility-group proteins from calf thymus, HMG-14 and HMG-20 (ubiquitin), and their interaction with DNA. Eur J Biochem., 112, 577-580. Castellucci M, Kaufmann P, Bischof P. (1990) Extracellular matrix influences hormone and protein production by human chorionic villi. Cell Tissue Res., 262, 135-142. Chang CW, Chuang HC, Yu C, Yao TP, Chen H. (2005) Stimulation of GCMa transcriptional activity by cyclic AMP/protein kinase A signaling is attributed to CBP-mediated acetylation of GCMa. Mol Cell Biol., 25, 8401-8414. Chen CP, Chen CY, Yang YC, Su TH, Chen H. (2004) Decreased placental GCM1 (glial cells missing) gene expression in pre-eclampsia. Placenta. 25, 413-421. Cohen SX, Moulin M, Schilling O, Meyer-Klaucke W, Schreiber J, Wegner M, Müller CW. (2002) The GCM domain is a Zn-coordinating DNA-binding domain. FEBS Lett., 528, 95-100 Cohen SX, Moulin M, Hashemolhosseini S, Kilian K, Wegner M, Müller CW. (2003) Structure of the GCM domain-DNA complex: a DNA-binding domain with a novel fold and mode of target site recognition. EMBO J., 22, 35-45. Coux O, Tanaka K, Goldberg AL. (1996) Structure and functions of the 20S and 26S proteasomes. Annu Rev Biochem., 65, 801-847. Deveraux Q, Jensen C, Rechsteiner M. (1995) Molecular cloning and expression of a 26 S protease subunit enriched in dileucine repeats. J Biol Chem., 270, 23726-23729. Deveraux Q, Ustrell V, Pickart C, Rechsteiner M. (1994) A 26 S protease subunit that binds ubiquitin conjugates. J Biol Chem., 269, 7059-7061. Dominguez C, Bonvin AM, Winkler GS, van Schaik FM, Timmers HT, Boelens R. (2004) Structural model of the UbcH5B/CNOT4 complex revealed by combining NMR, mutagenesis, and docking approaches. Structure., 12, 633-644. Furukawa M, Ohta T, Xiong Y. (2002) Activation of UBC5 ubiquitin-conjugating enzyme by the RING finger of ROC1 and assembly of active ubiquitin ligases by all cullins. J Biol Chem., 277, 15758-15765. Groll M, Bajorek M, Köhler A, Moroder L, Rubin DM, Huber R, Glickman MH, Finley D. (2000) A gated channel into the proteasome core particle. Nat Struct Biol., 7, 1062-1067. Haas AL, Rose IA. (1982) The mechanism of ubiquitin activating enzyme. A kinetic and equilibrium analysis. J Biol Chem., 257, 10329-10337. Hashemolhosseini S, Hadjihannas M, Stolt CC, Haas CS, Amann K, Wegner M. (2002) Restricted expression of mouse GCMa/Gcm1 in kidney and thymus. Mech Dev., 118, 175-178. Hashemolhosseini S, Kilian K, Kardash E, Lischka P, Stamminger T, Wegner M. (2003) Structural requirements for nuclear localization of GCMa/Gcm-1. FEBS Lett., 553, 315-320. Hashemolhosseini S, Schmidt K, Kilian K, Rodriguez E, Wegner M. (2004) Conservation and variation of structure and function in a newly identified GCM homolog from chicken. J Mol Biol., 336, 441-451. Hashemolhosseini S, Wegner M. (2004) Impacts of a new transcription factor family: mammalian GCM proteins in health and disease. J Cell Biol., 166, 765-768. Hershko A, Heller H, Elias S, Ciechanover A. (1983) Components of ubiquitin-protein ligase system. Resolution, affinity purification, and role in protein breakdown. J Biol Chem., 258, 8206-8214. Hochstrasser M. (1996) Ubiquitin-dependent protein degradation. Annu Rev Genet. 30, 405-439. Hosoya T, Takizawa K, Nitta K, Hotta Y. (1995) glial cells missing: a binary switch between neuronal and glial determination in Drosophila. Cell. 82, 1025-1036. Houben K, Dominguez C, van Schaik FM, Timmers HT, Bonvin AM, Boelens R. (2004) Solution structure of the ubiquitin-conjugating enzyme UbcH5B. J Mol Biol., 344, 513-526. Hough R, Pratt G, Rechsteiner M. (1986) Ubiquitin-lysozyme conjugates. Identification and characterization of an ATP-dependent protease from rabbit reticulocyte lysates. J Biol Chem., 261, 2400-2408. Hough R, Rechsteiner M. (1986) Ubiquitin-lysozyme conjugates. Purification and susceptibility to proteolysis. J Biol Chem., 261, 2391-2399. Huang DT, Hunt HW, Zhuang M, Ohi MD, Holton JM, Schulman BA. (2007) Basis for a ubiquitin-like protein thioester switch toggling E1-E2 affinity. Nature., 445, 394-398. Iwasaki Y, Hosoya T, Takebayashi H, Ogawa Y, Hotta Y, Ikenaka K. (2003) The potential to induce glial differentiation is conserved between Drosophila and mammalian glial cells missing genes. Development., 130, 6027-6035. Jensen JP, Bates PW, Yang M, Vierstra RD, Weissman AM. (1995) Identification of a family of closely related human ubiquitin conjugating enzymes. J Biol Chem., 270, 30408-30414. Jentsch S. (1992) The ubiquitin-conjugation system. Annu Rev Genet., 26, 179-207. Jentsch S. (1992) Ubiquitin-dependent protein degradation: a cellular perspective. Trends Cell Biol., 2, 98-103. Kammerer M, Giangrande A. Glide2, a second glial promoting factor in Drosophila melanogaster. EMBO J., 20, 4664-4673. Kim J, Jones BW, Zock C, Chen Z, Wang H, Goodman CS, Anderson DJ. (1998) Isolation and characterization of mammalian homologs of the Drosophila gene glial cells missing. Proc Natl Acad Sci U S A., 95, 12364-12369. Lois LM, Lima CD. (2005) Structures of the SUMO E1 provide mechanistic insights into SUMO activation and E2 recruitment to E1. EMBO J., 24, 439-451. Lorick KL, Jensen JP, Fang S, Ong AM, Hatakeyama S, Weissman AM. (1999) RING fingers mediate ubiquitin-conjugating enzyme (E2)-dependent ubiquitination. Proc Natl Acad Sci U S A., 96, 11364-11369. Mason GG, Murray RZ, Pappin D, Rivett AJ. (1998) Phosphorylation of ATPase subunits of the 26S proteasome. FEBS Lett., 430, 269-274. Miura T, Klaus W, Gsell B, Miyamoto C, Senn H. (1999) Characterization of the binding interface between ubiquitin and class I human ubiquitin-conjugating enzyme 2b by multidimensional heteronuclear NMR spectroscopy in solution. J Mol Biol., 290, 213-228. Nait-Oumesmar B, Copperman AB, Lazzarini RA. (2000) Placental expression and chromosomal localization of the human Gcm 1 gene. J Histochem Cytochem., 48, 915-922. Nait-Oumesmar B, Stecca B, Fatterpekar G, Naidich T, Corbin J, Lazzarini RA. (2002) Ectopic expression of Gcm1 induces congenital spinal cord abnormalities. Development., 129, 3957-3964. Pfrieger FW, Barres BA. (1996) New views on synapse-glia interactions. Curr Opin Neurobiol., 6, 615-621. Reifegerste R, Schreiber J, Gülland S, Lüdemann A, Wegner M. (1999) mGCMa is a murine transcription factor that overrides cell fate decisions in Drosophila. Mech Dev., 82, 141-150. Rubin DM, Finley D. (1995) Proteolysis. The proteasome: a protein-degrading organelle? Curr Biol., 5, 854-858. Schreiber J, Enderich J, Wegner M. (1998) Structural requirements for DNA binding of GCM proteins. Nucleic Acids Res., 26, 2337-2343. Seufert W, Jentsch S. (1992) In vivo function of the proteasome in the ubiquitin pathway. EMBO J., 11, 3077-3080. Tuerk EE, Schreiber J, Wegner M. (2000) Protein stability and domain topology determine the transcriptional activity of the mammalian glial cells missing homolog, GCMb. J Biol Chem., 275, 4774-4782. Wegner M, Riethmacher D. (2001) Chronicles of a switch hunt: gcm genes in development. Trends Genet., 17, 286-290. Yang CS, Yu C, Chuang HC, Chang CW, Chang GD, Yao TP, Chen H. (2005) FBW2 targets GCMa to the ubiquitin-proteasome degradation system. J Biol Chem., 280, 10083-10090. Yu C, Shen K, Lin M, Chen P, Lin C, Chang GD, Chen H. (2002) GCMa regulates the syncytin-mediated trophoblastic fusion. J Biol Chem., 277, 50062-50068. Zheng N, Wang P, Jeffrey PD, Pavletich NP. (2000) Structure of a c-Cbl-UbcH7 complex: RING domain function in ubiquitin-protein ligases. Cell., 102, 533-539. Zheng N, Schulman BA, Song L, Miller JJ, Jeffrey PD, Wang P, Chu C, Koepp DM, Elledge SJ, Pagano M, Conaway RC, Conaway JW, Harper JW, Pavletich NP. (2002) Structure of the Cul1-Rbx1-Skp1-F boxSkp2 SCF ubiquitin ligase complex. Nature., 416, 703-709. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/26523 | - |
dc.description.abstract | GCM1是在胎盤發育中一個必需的轉錄因子,而且它的活性已經被證實受到汎素-蛋白酶體的降解系統所調控。蛋白質的汎素化需要三種酵素的參與,分別是E1活化酵素、E2結合酵素以及E3接合酵素。在我們之前的研究中指出,SCFFBXW2複合體能夠藉由人類F-box蛋白FBW2專一性地辨識GCM1並扮演其E3接合酶的角色;然而,是哪一個E2結合酵素參與在其中仍然是個待解的習題。
在這個研究中,我們首先指出HeLa S-100萃取液可以在活體外促進GCM1的汎素化;並且進一步地在一系列的E2中鑑定出一個可以專一地調控GCM1汎素化的E2蛋白(稱之為E2-6)。我們也利用活體外汎素化的實驗證實了E1、E2-6和SCFFBXW2對於GCM1的汎素化都是不可或缺的。E2-6活性區的點突變會使得它失去和汎素結合的能力,在我們的實驗中也證實了這樣的突變會導致E2-6失去和E3的交互作用而無法將GCM1汎素化。另外,在293T細胞中利用RNA干擾抑制E2-6蛋白的表現會使得GCM1較不易被汎素化而能夠被保存下來。 上述的實驗結果顯示,無論在活體內或是活體外,E2-6都在GCM1的汎素化中扮演著重要角色。 | zh_TW |
dc.description.abstract | Protein ubiquitination involves E1 ubiquitin-activating enzyme, E2 ubiquitin-conjugating enzyme, and E3 ligase. GCM1, whose activity has been demonstrated to be regulated by the ubiquitin-proteasome system, is an essential transcription factor in placental development. Our previous studies have shown that GCM1 is a substrate for the SCFFBXW2 E3 complex, which specifically recognizes GCM1 via the human F-box protein, FBW2. However, the ubiquitin-conjugating enzyme (E2) involved in GCM1 ubiquitination remains elusive, in this study, we first showed that the HeLa S-100 extract can promote ubiquitination of GCM1 in vitro, and then identified an E2 (termed E2-6) specifically regulates GCM1 ubiquitination after screening a panel of E2s. In vitro ubiquitination assay of GCM1 also demonstrated that E1, E2-6, and SCFFBW2 are necessary for GCM1 ubiquitination. Mutation on the catalytic site of E2-6 (E2-6CA) resulted in loss of its interaction with SCFFBW2 and the ubiquitination of GCM1. Moreover, small hairpin RNA-mediated knockdown of E2-6 reduced ubiquitination of GCM1 and subsequently increased GCM1 protein stability in 293T cells. These results reveal that E2-6 is crucial for ubiquitination of GCM1 in vitro and in vivo. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T07:13:38Z (GMT). No. of bitstreams: 1 ntu-97-R94b46044-1.pdf: 1578875 bytes, checksum: 4507f0ab458960888286edfd963dfd37 (MD5) Previous issue date: 2008 | en |
dc.description.tableofcontents | 目錄……………………………………………………………… ……………………...I
圖表目錄……………………………………………………………………………….III 縮寫表………………………………………………………………………………….IV 中文摘要……………………………………………………………………………….VI 英文摘要Abstrate……………………………………………………………………..VII 第一章 序論………………………………………………………………………...1 1.1 胎盤………………………………………………………………………..…1 1.2 gcm轉錄因子基因家族……………………………………………………..3 1.3 GCMa的降解機制…………………………………………………………..5 1.4 E2結合酵素………………………………………………………………….7 1.5 研究動機……………………………………………………………………..8 第二章 材料及方法…………………………………………………………..9 2.1 重組質體的構築與增殖……………………………………………………..9 2.2 細胞培養……………………………………………………………………13 2.3 活體外汎素化反應分析…………………………………………………....15 2.4 西方墨點法…………………………………………………………………16 2.5 共同免疫沉澱法…………………………………………………………....16 2.6 GST融合蛋白表現………………………………………………………....17 2.7 活體內汎素化試驗………………………………………………………....17 2.8 35S脈衝追蹤試驗…………………………………………………………..18 2.9 反轉錄聚合酶連鎖反應…………………………………………………....18 第三章 實驗結果………………………………………………………………….20 3.1 汎素在活體外對GCM1的標定………………………………….………..20 3.2 E2-6可以專一性地促進GCM1汎素化…………………………………..20 3.3 E2-6在GCM1汎素化中扮演E2結合酵素的角色…………………….…21 3.4 E2-6和SCFFBW2複合體的交互作用……………………………………...22 3.5 利用外生性shRNA質體抑制細胞內E2-6蛋白表現………………….…23 3.6 E2-6對GCM1汎素化及蛋白穩定度的影響……………………………..23 第四章 討論與總結……………………………………………………………….25 第五章 圖表……………………………………………………………………….28 第六章 參考文獻………………………………………………………………….36 | |
dc.language.iso | zh-TW | |
dc.title | 鑑定參與GCM1汎素化之E2結合酵素及其特性探討 | zh_TW |
dc.title | Identification and Characterization of an E2 Ubiquitin-Conjugating Enzyme in GCM1 Ubiquitination | en |
dc.type | Thesis | |
dc.date.schoolyear | 96-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 黃詮珍,張震東,李明亭,張功耀 | |
dc.subject.keyword | 胎盤, GCM1, SCF複合體, FBXW2, E2-6, | zh_TW |
dc.subject.keyword | Placenta, GCM1, SCF complex, FBXW2, E2-6, | en |
dc.relation.page | 40 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2008-07-30 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 生化科學研究所 | zh_TW |
顯示於系所單位: | 生化科學研究所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-97-1.pdf 目前未授權公開取用 | 1.54 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。