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作者(中文):陳俊豪
作者(外文):CHEN, CHUN-HAO
論文名稱(中文):金屬感應轉錄因子與缺氧誘導因子之交互作用及對其轉錄活性的影響
論文名稱(外文):MTF-1 interacts with HIF-1α and regulates its transcriptional activity
指導教授(中文):林立元
口試委員(中文):李易展
楊嘉鈴
學位類別:碩士
校院名稱:國立清華大學
系所名稱:分子與細胞生物研究所
學號:100080540
出版年(民國):102
畢業學年度:102
語文別:中文
論文頁數:63
中文關鍵詞:金屬感應轉錄因子缺氧誘導因子
外文關鍵詞:MTF-1HIF-1αZinc
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缺氧誘導因子HIF-1α是腫瘤發展初期很重要的指標性蛋白之一,其會受到腫瘤內部產生之缺氧環境誘導表現且促使血管新生導致腫瘤轉移。金屬感應轉錄因子MTF-1在哺乳類動物胚胎發育扮演一個重要的角色,控制了多數與細胞內鋅離子恆定及抗氧化之相關基因的表現。本篇研究中我們首先利用共同免疫沉澱技術證實了在細胞內兩者蛋白有交互作用的現象發生,利用MTF-1不同片段的質體與HIF-1α共同表現,發現HIF-1α與MTF-1之所有區域皆發生交互作用的現象,且利用HIF-1α之去除ODD區域質體,我們也證實HIF-1α負責蛋白質穩定度之區域並非與MTF-1交互作用主要片段。另外本篇研究發現細胞在鋅離子的刺激下,MTF-1與HIF-1α交互作用會增高,且該交互作用會隨著鋅離子處理劑量逐漸升高而加強,推測可能與HIF-1α受到鋅離子刺激有關,透過西方墨點法我們證實施加鋅離子會導致HIF-1α蛋白表現增加,且利用cycloheximide的處理發現此表現量增加並非透過增強蛋白質穩定度的途徑。接下來我們利用分離細胞質與細胞核萃取液的方法探討在MTF-1存在是否會影響HIF-1α的細胞質與細胞核的分佈,研究結果顯示MTF-1的存在並沒有顯著影響HIF-1α的分佈,然而利用報導基因活性檢測技術及siRNA降低基因表現程度發現MTF-1的存在使HIF-1α轉錄活性顯著提升。
Hypoxia-inducible factor- 1α (HIF-1α) is an important protein involved in tumor development. Production of HIF-1α would be stimulated by the hypoxic environment in tumor and leads to tumor metastasis. Metal responsive transcription factor 1 (MTF-1), which plays an essential role in modulating the expression of genes, involves in cellular zinc homeostasis. In this study, we investigated the interaction between HIF-1α and MTF-1 by immunoprecipitation. The results revealed that HIF-1α interacts with all of the domains of MTF-1. Besides, we also expressed MTF-1 and HIF-1α with the deletion of ODD domain. The results showed that loss of ODD domain, a region responsible for protein stability, had no effect on HIF-1α and MTF-1 interaction. Noticeably, addition of zinc to cells increased MTF-1/ HIF-1α interaction in a dose-dependent manner. According to the results, addition of zinc increased the HIF-1α protein level. However, this increase was not due to the alternation of its mRNA level by enhancing protein stability through the use of cycloheximide. Analyzing the position of HIF-1α in cytoplasmic and nuclear fraction showed that the presence of MTF-1 did not affect the distribution of HIF-1α. Using reporter gene assay and siRNA to reduce the expression of MTF-1 mRNA revealed that the presence of MTF-1 would increase the transcriptional activity of HIF-1α significantly.
目錄
中文摘要 2
英文摘要 3
縮寫表 4
前言 5
材料與方法 15
結果 24
討論 31
參考資料 39
附圖 46
參考文獻
Andrews GK, Lee DK, Ravindra R, Lichtlen P, Sirito M, Sawadogo M, Schaffner W (2001) The transcription factors MTF-1 and USF1 cooperate to regulate mouse metallothionein-I expression in response to the essential metal zinc in visceral endoderm cells during early development. EMBO J 20(5): 1114-1122

Bae MK, Ahn MY, Jeong JW, Bae MH, Lee YM, Bae SK, Park JW, Kim KR, Kim KW (2002) Jab1 interacts directly with HIF-1alpha and regulates its stability. J Biol Chem 277(1): 9-12

Blum R, Jacob-Hirsch J, Amariglio N, Rechavi G, Kloog Y (2005) Ras inhibition in glioblastoma down-regulates hypoxia-inducible factor-1alpha, causing glycolysis shutdown and cell death. Cancer research 65(3): 999-1006

Brugnera E, Georgiev O, Radtke F, Heuchel R, Baker E, Sutherland GR, Schaffner W (1994) Cloning, chromosomal mapping and characterization of the human metal-regulatory transcription factor MTF-1. Nucleic Acids Res 22(15): 3167-3173

Cheng J, Kang X, Zhang S, Yeh ET (2007) SUMO-specific protease 1 is essential for stabilization of HIF1alpha during hypoxia. Cell 131(3): 584-595

Compernolle V, Brusselmans K, Acker T, Hoet P, Tjwa M, Beck H, Plaisance S, Dor Y, Keshet E, Lupu F, Nemery B, Dewerchin M, Van Veldhoven P, Plate K, Moons L, Collen D, Carmeliet P (2002) Loss of HIF-2alpha and inhibition of VEGF impair fetal lung maturation, whereas treatment with VEGF prevents fatal respiratory distress in premature mice. Nature medicine 8(7): 702-710

Cramer M, Nagy I, Murphy BJ, Gassmann M, Hottiger MO, Georgiev O, Schaffner W (2005) NF-kappaB contributes to transcription of placenta growth factor and interacts with metal responsive transcription factor-1 in hypoxic human cells. Biol Chem 386(9): 865-872

Dai S, Yin Z, Yuan G, Lu H, Jia R, Xu J, Song X, Li L, Shu Y, Liang X, He C, Lv C, Zhang W (2013) Quantification of metallothionein on the liver and kidney of rats by subchronic lead and cadmium in combination. Environmental toxicology and pharmacology 36(3): 1207-1216

Datta J, Majumder S, Kutay H, Motiwala T, Frankel W, Costa R, Cha HC, MacDougald OA, Jacob ST, Ghoshal K (2007) Metallothionein expression is suppressed in primary human hepatocellular carcinomas and is mediated through inactivation of CCAAT/enhancer binding protein alpha by phosphatidylinositol 3-kinase signaling cascade. Cancer research 67(6): 2736-2746

Dube A, Harrisson JF, Saint-Gelais G, Seguin C Hypoxia acts through multiple signaling pathways to induce metallothionein transactivation by the metal-responsive transcription factor-1 (MTF-1). Biochem Cell Biol 89(6): 562-577

Dunn MF (2005) Zinc-ligand interactions modulate assembly and stability of the insulin hexamer -- a review. Biometals : an international journal on the role of metal ions in biology, biochemistry, and medicine 18(4): 295-303

Duyndam MC, Hulscher TM, Fontijn D, Pinedo HM, Boven E (2001) Induction of vascular endothelial growth factor expression and hypoxia-inducible factor 1alpha protein by the oxidative stressor arsenite. J Biol Chem 276(51): 48066-48076

Fels DR, Koumenis C (2005) HIF-1alpha and p53: the ODD couple? Trends Biochem Sci 30(8): 426-429

Fujimoto S, Itsumura N, Tsuji T, Anan Y, Tsuji N, Ogra Y, Kimura T, Miyamae Y, Masuda S, Nagao M, Kambe T (2013) Cooperative Functions of ZnT1, Metallothionein and ZnT4 in the Cytoplasm Are Required for Full Activation of TNAP in the Early Secretory Pathway. PloS one 8(10): e77445

Gao N, Ding M, Zheng JZ, Zhang Z, Leonard SS, Liu KJ, Shi X, Jiang BH (2002) Vanadate-induced expression of hypoxia-inducible factor 1 alpha and vascular endothelial growth factor through phosphatidylinositol 3-kinase/Akt pathway and reactive oxygen species. J Biol Chem 277(35): 31963-31971

Gray MJ, Zhang J, Ellis LM, Semenza GL, Evans DB, Watowich SS, Gallick GE (2005) HIF-1alpha, STAT3, CBP/p300 and Ref-1/APE are components of a transcriptional complex that regulates Src-dependent hypoxia-induced expression of VEGF in pancreatic and prostate carcinomas. Oncogene 24(19): 3110-3120

Green CJ, Lichtlen P, Huynh NT, Yanovsky M, Laderoute KR, Schaffner W, Murphy BJ (2001) Placenta growth factor gene expression is induced by hypoxia in fibroblasts: a central role for metal transcription factor-1. Cancer research 61(6): 2696-2703

Guerrerio AL, Berg JM (2004) Metal ion affinities of the zinc finger domains of the metal responsive element-binding transcription factor-1 (MTF1). Biochemistry 43(18): 5437-5444

Gunther V, Lindert U, Schaffner W The taste of heavy metals: gene regulation by MTF-1. Biochim Biophys Acta 1823(9): 1416-1425

Isaacs JS, Jung YJ, Mimnaugh EG, Martinez A, Cuttitta F, Neckers LM (2002) Hsp90 regulates a von Hippel Lindau-independent hypoxia-inducible factor-1 alpha-degradative pathway. J Biol Chem 277(33): 29936-29944

Ivan M, Haberberger T, Gervasi DC, Michelson KS, Gunzler V, Kondo K, Yang H, Sorokina I, Conaway RC, Conaway JW, Kaelin WG, Jr. (2002) Biochemical purification and pharmacological inhibition of a mammalian prolyl hydroxylase acting on hypoxia-inducible factor. Proc Natl Acad Sci U S A 99(21): 13459-13464

Joshi JG, Zimmerman A (1988) Ferritin: an expanded role in metabolic regulation. Toxicology 48(1): 21-29

Ke Q, Costa M (2006) Hypoxia-inducible factor-1 (HIF-1). Mol Pharmacol 70(5): 1469-1480

Kimura T, Li Y, Okumura F, Itoh N, Nakanishi T, Sone T, Isobe M, Andrews GK (2008) Chromium(VI) inhibits mouse metallothionein-I gene transcription by preventing the zinc-dependent formation of an MTF-1-p300 complex. Biochem J 415(3): 477-482

Kitajima Y, Miyazaki K (2013) The Critical Impact of HIF-1a on Gastric Cancer Biology. Cancers 5(1): 15-26

LaRochelle O, Labbe S, Harrisson JF, Simard C, Tremblay V, St-Gelais G, Govindan MV, Seguin C (2008) Nuclear factor-1 and metal transcription factor-1 synergistically activate the mouse metallothionein-1 gene in response to metal ions. J Biol Chem 283(13): 8190-8201

Lee SH, Bae SC, Kim KW, Lee YM RUNX3 inhibits hypoxia-inducible factor-1alpha protein stability by interacting with prolyl hydroxylases in gastric cancer cells. Oncogene

Li Y, Kimura T, Huyck RW, Laity JH, Andrews GK (2008) Zinc-induced formation of a coactivator complex containing the zinc-sensing transcription factor MTF-1, p300/CBP, and Sp1. Mol Cell Biol 28(13): 4275-4284

Lichtlen P, Schaffner W (2001) The "metal transcription factor" MTF-1: biological facts and medical implications. Swiss Med Wkly 131(45-46): 647-652

Lin CY, Liu YC, Lin MC, Thi Nguyen T, Tam MF, Chein CY, Lin MT, Lin LY (2013) Expression and characterization of SUMO-conjugated metal-responsive transcription factor 1: SIM-dependent cross-interaction and distinct DNA binding activity. Journal of biochemistry 153(4): 361-369

Lin MC, Liu YC, Tam MF, Lu YJ, Hsieh YT, Lin LY PTEN interacts with metal-responsive transcription factor 1 and stimulates its transcriptional activity. Biochem J 441(1): 367-377

Majumder S, Ghoshal K, Gronostajski RM, Jacob ST (2001) Downregulation of constitutive and heavy metal-induced metallothionein-I expression by nuclear factor I. Gene Expr 9(4-5): 203-215

Makino Y, Cao R, Svensson K, Bertilsson G, Asman M, Tanaka H, Cao Y, Berkenstam A, Poellinger L (2001) Inhibitory PAS domain protein is a negative regulator of hypoxia-inducible gene expression. Nature 414(6863): 550-554

Migdal M, Huppertz B, Tessler S, Comforti A, Shibuya M, Reich R, Baumann H, Neufeld G (1998) Neuropilin-1 is a placenta growth factor-2 receptor. J Biol Chem 273(35): 22272-22278

Moroz E, Carlin S, Dyomina K, Burke S, Thaler HT, Blasberg R, Serganova I (2009) Real-time imaging of HIF-1alpha stabilization and degradation. PloS one 4(4): e5077

Murphy BJ, Andrews GK, Bittel D, Discher DJ, McCue J, Green CJ, Yanovsky M, Giaccia A, Sutherland RM, Laderoute KR, Webster KA (1999) Activation of metallothionein gene expression by hypoxia involves metal response elements and metal transcription factor-1. Cancer research 59(6): 1315-1322

Murphy BJ, Kimura T, Sato BG, Shi Y, Andrews GK (2008) Metallothionein induction by hypoxia involves cooperative interactions between metal-responsive transcription factor-1 and hypoxia-inducible transcription factor-1alpha. Mol Cancer Res 6(3): 483-490

Murphy BJ, Sato BG, Dalton TP, Laderoute KR (2005) The metal-responsive transcription factor-1 contributes to HIF-1 activation during hypoxic stress. Biochem Biophys Res Commun 337(3): 860-867

Nardinocchi L, Pantisano V, Puca R, Porru M, Aiello A, Grasselli A, Leonetti C, Safran M, Rechavi G, Givol D, Farsetti A, D'Orazi G Zinc downregulates HIF-1alpha and inhibits its activity in tumor cells in vitro and in vivo. PloS one 5(12): e15048

Nieminen AL, Qanungo S, Schneider EA, Jiang BH, Agani FH (2005) Mdm2 and HIF-1alpha interaction in tumor cells during hypoxia. J Cell Physiol 204(2): 364-369

Okumura F, Li Y, Itoh N, Nakanishi T, Isobe M, Andrews GK, Kimura T The zinc-sensing transcription factor MTF-1 mediates zinc-induced epigenetic changes in chromatin of the mouse metallothionein-I promoter. Biochim Biophys Acta 1809(1): 56-62

Park SE, Park JW, Cho YS, Ryu JH, Paick JS, Chun YS (2007) HIF-1alpha promotes survival of prostate cells at a high zinc environment. Prostate 67(14): 1514-1523

Poon E, Harris AL, Ashcroft M (2009) Targeting the hypoxia-inducible factor (HIF) pathway in cancer. Expert reviews in molecular medicine 11: e26

Powis G, Kirkpatrick L (2004) Hypoxia inducible factor-1alpha as a cancer drug target. Molecular cancer therapeutics 3(5): 647-654

Ratcliffe PJ, O'Rourke JF, Maxwell PH, Pugh CW (1998) Oxygen sensing, hypoxia-inducible factor-1 and the regulation of mammalian gene expression. J Exp Biol 201(Pt 8): 1153-1162

Salnikow K, Su W, Blagosklonny MV, Costa M (2000) Carcinogenic metals induce hypoxia-inducible factor-stimulated transcription by reactive oxygen species-independent mechanism. Cancer research 60(13): 3375-3378

Semenza GL, Nejfelt MK, Chi SM, Antonarakis SE (1991) Hypoxia-inducible nuclear factors bind to an enhancer element located 3' to the human erythropoietin gene. Proc Natl Acad Sci U S A 88(13): 5680-5684

Shi Y, Amin K, Sato BG, Samuelsson SJ, Sambucetti L, Haroon ZA, Laderoute K, Murphy BJ The metal-responsive transcription factor-1 protein is elevated in human tumors. Cancer Biol Ther 9(6): 469-476

Troadec MB, Ward DM, Lo E, Kaplan J, De Domenico I Induction of FPN1 transcription by MTF-1 reveals a role for ferroportin in transition metal efflux. Blood 116(22): 4657-4664

Uenishi R, Gong P, Suzuki K, Koizumi S (2006) Cross talk of heat shock and heavy metal regulatory pathways. Biochem Biophys Res Commun 341(4): 1072-1077

Vaupel P, Harrison L (2004) Tumor hypoxia: causative factors, compensatory mechanisms, and cellular response. Oncologist 9 Suppl 5: 4-9

Wan J, Chai H, Yu Z, Ge W, Kang N, Xia W, Che Y HIF-1alpha effects on angiogenic potential in human small cell lung carcinoma. J Exp Clin Cancer Res 30: 77

Welsh SJ, Bellamy WT, Briehl MM, Powis G (2002) The redox protein thioredoxin-1 (Trx-1) increases hypoxia-inducible factor 1alpha protein expression: Trx-1 overexpression results in increased vascular endothelial growth factor production and enhanced tumor angiogenesis. Cancer research 62(17): 5089-5095

Wiesener MS, Turley H, Allen WE, Willam C, Eckardt KU, Talks KL, Wood SM, Gatter KC, Harris AL, Pugh CW, Ratcliffe PJ, Maxwell PH (1998) Induction of endothelial PAS domain protein-1 by hypoxia: characterization and comparison with hypoxia-inducible factor-1alpha. Blood 92(7): 2260-2268

Wu D, Zhang R, Zhao R, Chen G, Cai Y, Jin J A novel function of novobiocin: disrupting the interaction of HIF 1alpha and p300/CBP through direct binding to the HIF1alpha C-terminal activation domain. PloS one 8(5): e62014

Yoo YG, Kong G, Lee MO (2006) Metastasis-associated protein 1 enhances stability of hypoxia-inducible factor-1alpha protein by recruiting histone deacetylase 1. EMBO J 25(6): 1231-1241

Zundel W, Schindler C, Haas-Kogan D, Koong A, Kaper F, Chen E, Gottschalk AR, Ryan HE, Johnson RS, Jefferson AB, Stokoe D, Giaccia AJ (2000) Loss of PTEN facilitates HIF-1-mediated gene expression. Genes Dev 14(4): 391-396


 
 
 
 
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