臺灣博碩士論文加值系統

中文摘要
基因要轉錄出mRNA必須透過RNA polymerase II的作用，而TATA-less啟動子的轉錄起始已知是由TFIID與Sp1結合在GC box序列而引發的，其中TFIID是一個多分子組成的複合體，其組成包含了TAFII250, TAFII150及TAFII130與結合在啟動子GC區域上的Sp1。EB病毒 (Epstein-Barr virus) 含有兩個生活史，當病毒感染B淋巴細胞之後通常以潛伏型態存在，然而，EB病毒的增生必須透過溶裂循環，在宿主細胞受到一些特定的化學或是物理刺激後，能刺激病毒引發溶裂循環，當EB病毒進入溶裂循環，EB病毒會表現Rta及Zta兩種轉錄因子，分別是由BRLF1與BZLF1基因所轉譯出來。目前已知Rta不僅會結合到啟動子上的RRE序列來活化基因轉錄，也會間接的與Sp1結合，以共同活化子的身分來增進基因的轉錄活性。本篇研究目的主要是在探討Rta是否會藉由與TAFII130結合來活化缺乏TATA序列的啟動子的表現。首先利用DNA affinity precipitation assay與染色體免疫沉澱分析(chromatin immunoprecipitation analysis)發現Rta會與TAFII130及Sp1蛋白質形成複合體結合在EB病毒的LMP1基因與BALF5基因的TATA-less啟動子上，並更進一步的觀察到Rta與TAFII130間有交互作用，並會聚集在細胞核中，此外，結合區域分析也證實了Rta是透過TAFII130的C端部份與TAFII130結合，最後，Rta也可以活化LMP1、BALF5及人類androgen receptor等三個TATA-less啟動子的轉錄活性達9-17倍。綜合以上結論，這個研究證實了Rta可以透過與TAFII130與Sp1的結合來活化TATA-less啟動子的表現。

Abstract
In eukaryotes, gene encoding mRNA are transcribed by RNA polymerase II, a multiprotein enzyme. Initiation of TATA-less promotes has been involved in binding of the TFIID, including TAFII250, TAFII150, and TAFII110, with Sp1 bound at the GC boxes. Epstein-Barr virus (EBV) has two distinct life cycles. EBV is normally maintained under latent conditions after infecting B lymphocytes. However, EBV must enter a lytic cycle to proliferate. The lytic cycle can be activated in vitro by treating the cell with chemicals or physical stimulus. When EBV enters the lytic cycle, the virus expresses two transcription factors, Rta and Zta, which are encoded by BRLF1 and BZLF1, respectively. Rta not only binds to RRE in promoters to activate transcription but also indirectly interacts with Sp1 as a coactivator to enhance the transcription that depends on Sp1. The aim of this work is to demonstrate whether Rta activates TATA-less promoters via an interaction with TAFII130. By using DNA affinity precipitation assay and chromatin immunoprecipitation analysis, the Rta-TAFII130-Sp1 complex binds to the Sp1 sites on the TATA-less promoters of LMP1 and BALF5 of EBV. Moreover, Rta interacts and colocalizes with TAFII130 in the nucleus. In addition, binding domain analysis shows that Rta interacts with the C-terminal region of TAFII130. Finally, Rta transactivates three TATA-less promoters including LMP1、BALF5 and human androgen receptor by a factor 9-17. Taken together, this study reveals that Rta activates TATA-less promoters through an interaction with TAFII130 and Sp1.

中文摘要 P.4
英文摘要 P.5
前言 P.6
研究目的 P.16
材料方法 P.17
結果 P.23
討論 P.35
附表一 P.39
參考文獻 P.42

1.Adamson, A. L., D. Darr, E. Holley-Guthrie, R. A. Johnson, A. Mauser, J. Swenson, and S. Kenney. 2000. Epstein-Barr virus immediate-early proteins BZLF1 and BRLF1 activate the ATF2 transcription factor by increasing the levels of phosphorylated p38 and c-Jun N-terminal kinases. J Virol 74:1224-33.

2.Adamson, A. L., and S. Kenney. 1999. The Epstein-Barr virus BZLF1 protein interacts physically and functionally with the histone acetylase CREB-binding protein. J Virol 73:6551-8.

3.Amrolia, P. J., L. Ramamurthy, D. Saluja, N. Tanese, S. M. Jane, and J. M. Cunningham. 1997. The activation domain of the enhancer binding protein p45NF-E2 interacts with TAFII130 and mediates long-range activation of the alpha- and beta-globin gene loci in an erythroid cell line. Proc Natl Acad Sci U S A 94:10051-6.

4.Biggin, M., M. Bodescot, M. Perricaudet, and P. Farrell. 1987. Epstein-Barr virus gene expression in P3HR1-superinfected Raji cells. J Virol 61:3120-32.

5.Burley, S. K., and R. G. Roeder. 1996. Biochemistry and structural biology of transcription factor IID (TFIID). Annu Rev Biochem 65:769-99.

6.Chang, L. K., J. Y. Chung, Y. R. Hong, T. Ichimura, M. Nakao, and S. T. Liu. 2005. Activation of Sp1-mediated transcription by Rta of Epstein-Barr virus via an interaction with MCAF1. Nucleic Acids Res 33:6528-39.

7.Chang, L. K., and S. T. Liu. 2000. Activation of the BRLF1 promoter and lytic cycle of Epstein-Barr virus by histone acetylation. Nucleic Acids Res 28:3918-25.

8.Chang, Y., H. H. Lee, S. S. Chang, T. Y. Hsu, P. W. Wang, Y. S. Chang, K. Takada, and C. H. Tsai. 2004. Induction of Epstein-Barr virus latent membrane protein 1 by a lytic transactivator Rta. J Virol 78:13028-36.


9.Chen, J. L., L. D. Attardi, C. P. Verrijzer, K. Yokomori, and R. Tjian. 1994. Assembly of recombinant TFIID reveals differential coactivator requirements for distinct transcriptional activators. Cell 79:93-105.

10.Chevallier-Greco, A., H. Gruffat, E. Manet, A. Calender, and A. Sergeant. 1989. The Epstein-Barr virus (EBV) DR enhancer contains two functionally different domains: domain A is constitutive and cell specific, domain B is transactivated by the EBV early protein R. J Virol 63:615-23.

11.Chiang, C. M., and R. G. Roeder. 1995. Cloning of an intrinsic human TFIID subunit that interacts with multiple transcriptional activators. Science 267:531-6.

12.Cohen, J. I., F. Wang, J. Mannick, and E. Kieff. 1989. Epstein-Barr virus nuclear protein 2 is a key determinant of lymphocyte transformation. Proc Natl Acad Sci U S A 86:9558-62.

13.Cox, M. A., J. Leahy, and J. M. Hardwick. 1990. An enhancer within the divergent promoter of Epstein-Barr virus responds synergistically to the R and Z transactivators. J Virol 64:313-21.

14.Dambaugh, T., K. Hennessy, L. Chamnankit, and E. Kieff. 1984. U2 region of Epstein-Barr virus DNA may encode Epstein-Barr nuclear antigen 2. Proc Natl Acad Sci U S A 81:7632-6.

15.Dolyniuk, M., R. Pritchett, and E. Kieff. 1976. Proteins of Epstein-Barr virus. I. Analysis of the polypeptides of purified enveloped Epstein-Barr virus. J Virol 17:935-49.

16.Dolyniuk, M., E. Wolff, and E. Kieff. 1976. Proteins of Epstein-Barr Virus. II. Electrophoretic analysis of the polypeptides of the nucleocapsid and the glucosamine- and polysaccharide-containing components of enveloped virus. J Virol 18:289-97.

17.Epstein, M. A., and Y. M. Barr. 1964. Cultivation in Vitro of Human Lymphoblasts from Burkitt''s Malignant Lymphoma. Lancet 41:252-3.


18.Flemington, E., and S. H. Speck. 1990. Identification of phorbol ester response elements in the promoter of Epstein-Barr virus putative lytic switch gene BZLF1. J Virol 64:1217-26.

19.Flemington, E. K., A. E. Goldfeld, and S. H. Speck. 1991. Efficient transcription of the Epstein-Barr virus immediate-early BZLF1 and BRLF1 genes requires protein synthesis. J Virol 65:7073-7.

20.Fujita, N., S. Watanabe, T. Ichimura, Y. Ohkuma, T. Chiba, H. Saya, and M. Nakao. 2003. MCAF mediates MBD1-dependent transcriptional repression. Mol Cell Biol 23:2834-43.

21.Geser, A., G. de The, G. Lenoir, N. E. Day, and E. H. Williams. 1982. Final case reporting from the Ugandan prospective study of the relationship between EBV and Burkitt''s lymphoma. Int J Cancer 29:397-400.

22.Giot, J. F., I. Mikaelian, M. Buisson, E. Manet, I. Joab, J. C. Nicolas, and A. Sergeant. 1991. Transcriptional interference between the EBV transcription factors EB1 and R: both DNA-binding and activation domains of EB1 are required. Nucleic Acids Res 19:1251-8.

23.Given, D., and E. Kieff. 1979. DNA of Epstein-Barr virus. VI. Mapping of the internal tandem reiteration. J Virol 31:315-24.

24.Gruffat, H., N. Duran, M. Buisson, F. Wild, R. Buckland, and A. Sergeant. 1992. Characterization of an R-binding site mediating the R-induced activation of the Epstein-Barr virus BMLF1 promoter. J Virol 66:46-52.

25.Gruffat, H., E. Manet, A. Rigolet, and A. Sergeant. 1990. The enhancer factor R of Epstein-Barr virus (EBV) is a sequence-specific DNA binding protein. Nucleic Acids Res 18:6835-43.

26.Gruffat, H., and A. Sergeant. 1994. Characterization of the DNA-binding site repertoire for the Epstein-Barr virus transcription factor R. Nucleic Acids Res 22:1172-8.

27.Hammerschmidt, W., and B. Sugden. 1989. Genetic analysis of immortalizing functions of Epstein-Barr virus in human B lymphocytes. Nature 340:393-7.
28.Heller, M., P. Gerber, and E. Kieff. 1982. DNA of herpesvirus pan, a third member of the Epstein-Barr virus-Herpesvirus papio group. J Virol 41:931-9.

29.Henderson, E. E., and W. K. Long. 1981. Host cell reactivation of uv- and X-ray-damaged herpes simplex virus by Epstein-Barr virus (EBV)-transformed lymphoblastoid cell lines. Virology 115:237-48.

30.Henle, W., V. Diehl, G. Kohn, H. Zur Hausen, and G. Henle. 1967. Herpes-type virus and chromosome marker in normal leukocytes after growth with irradiated Burkitt cells. Science 157:1064-5.

31.Henle, W., K. Hummeler, and G. Henle. 1966. Antibody coating and agglutination of virus particles separated from the EB3 line of Burkitt lymphoma cells. J Bacteriol 92:269-71.

32.Hennessy, K., S. Fennewald, M. Hummel, T. Cole, and E. Kieff. 1984. A membrane protein encoded by Epstein-Barr virus in latent growth-transforming infection. Proc Natl Acad Sci U S A 81:7207-11.

33.Hennessy, K., S. Fennewald, and E. Kieff. 1985. A third viral nuclear protein in lymphoblasts immortalized by Epstein-Barr virus. Proc Natl Acad Sci U S A 82:5944-8.

34.Hennessy, K., and E. Kieff. 1983. One of two Epstein-Barr virus nuclear antigens contains a glycine-alanine copolymer domain. Proc Natl Acad Sci U S A 80:5665-9.

35.Hernandez, N. 1993. TBP, a universal eukaryotic transcription factor? Genes Dev 7:1291-308.

36.Hoey, T., R. O. Weinzierl, G. Gill, J. L. Chen, B. D. Dynlacht, and R. Tjian. 1993. Molecular cloning and functional analysis of Drosophila TAF110 reveal properties expected of coactivators. Cell 72:247-60.

37.Holley-Guthrie, E. A., E. B. Quinlivan, E. C. Mar, and S. Kenney. 1990. The Epstein-Barr virus (EBV) BMRF1 promoter for early antigen (EA-D) is regulated by the EBV transactivators, BRLF1 and BZLF1, in a cell-specific manner. J Virol 64:3753-9.
38.Hsu, T. Y., Y. Chang, P. W. Wang, M. Y. Liu, M. R. Chen, J. Y. Chen, and C. H. Tsai. 2005. Reactivation of Epstein-Barr virus can be triggered by an Rta protein mutated at the nuclear localization signal. J Gen Virol 86:317-22.

39.Jacq, X., C. Brou, Y. Lutz, I. Davidson, P. Chambon, and L. Tora. 1994. Human TAFII30 is present in a distinct TFIID complex and is required for transcriptional activation by the estrogen receptor. Cell 79:107-17.

40.Jones, J. F., S. Shurin, C. Abramowsky, R. R. Tubbs, C. G. Sciotto, R. Wahl, J. Sands, D. Gottman, B. Z. Katz, and J. Sklar. 1988. T-cell lymphomas containing Epstein-Barr viral DNA in patients with chronic Epstein-Barr virus infections. N Engl J Med 318:733-41.

41.Kaye, K. M., K. M. Izumi, and E. Kieff. 1993. Epstein-Barr virus latent membrane protein 1 is essential for B-lymphocyte growth transformation. Proc Natl Acad Sci U S A 90:9150-4.

42.Kenney, S. C., E. Holley-Guthrie, E. B. Quinlivan, D. Gutsch, Q. Zhang, T. Bender, J. F. Giot, and A. Sergeant. 1992. The cellular oncogene c-myb can interact synergistically with the Epstein-Barr virus BZLF1 transactivator in lymphoid cells. Mol Cell Biol 12:136-46.

43.Klein, G., J. Zeuthen, R. Ber, and I. Ernberg. 1982. Human lymphoma-lymphoma hybrids and lymphoma-leukemia hybrids. II. Epstein-Barr virus induction patterns. J Natl Cancer Inst 68:197-202.

44.Kollmar, R., and P. J. Farnham. 1993. Site-specific initiation of transcription by RNA polymerase II. Proc Soc Exp Biol Med 203:127-39.

45.Lee, M. A., M. E. Diamond, and J. L. Yates. 1999. Genetic evidence that EBNA-1 is needed for efficient, stable latent infection by Epstein-Barr virus. J Virol 73:2974-82.

46.Lieberman, P. M., and A. J. Berk. 1991. The Zta trans-activator protein stabilizes TFIID association with promoter DNA by direct protein-protein interaction. Genes Dev 5:2441-54.


47.Liu, C., N. D. Sista, and J. S. Pagano. 1996. Activation of the Epstein-Barr virus DNA polymerase promoter by the BRLF1 immediate-early protein is mediated through USF and E2F. J Virol 70:2545-55.

48.Longnecker, R., and E. Kieff. 1990. A second Epstein-Barr virus membrane protein (LMP2) is expressed in latent infection and colocalizes with LMP1. J Virol 64:2319-26.

49.Longnecker, R., C. L. Miller, B. Tomkinson, X. Q. Miao, and E. Kieff. 1993. Deletion of DNA encoding the first five transmembrane domains of Epstein-Barr virus latent membrane proteins 2A and 2B. J Virol 67:5068-74.

50.Maldonado, E., and D. Reinberg. 1995. News on initiation and elongation of transcription by RNA polymerase II. Curr Opin Cell Biol 7:352-61.

51.Manet, E., H. Gruffat, M. C. Trescol-Biemont, N. Moreno, P. Chambard, J. F. Giot, and A. Sergeant. 1989. Epstein-Barr virus bicistronic mRNAs generated by facultative splicing code for two transcriptional trans-activators. Embo J 8:1819-26.

52.Manet, E., A. Rigolet, H. Gruffat, J. F. Giot, and A. Sergeant. 1991. Domains of the Epstein-Barr virus (EBV) transcription factor R required for dimerization, DNA binding and activation. Nucleic Acids Res 19:2661-7.

53.Mannick, J. B., J. I. Cohen, M. Birkenbach, A. Marchini, and E. Kieff. 1991. The Epstein-Barr virus nuclear protein encoded by the leader of the EBNA RNAs is important in B-lymphocyte transformation. J Virol 65:6826-37.

54.Mueller, N., A. Evans, N. L. Harris, G. W. Comstock, E. Jellum, K. Magnus, N. Orentreich, B. F. Polk, and J. Vogelman. 1989. Hodgkin''s disease and Epstein-Barr virus. Altered antibody pattern before diagnosis. N Engl J Med 320:689-95.

55.Niedobitek, G., S. Hamilton-Dutoit, H. Herbst, T. Finn, M. Vetner, G. Pallesen, and H. Stein. 1989. Identification of Epstein-Barr virus-infected cells in tonsils of acute infectious mononucleosis by in situ hybridization. Hum Pathol 20:796-9.

56.Pugh, B. F., and R. Tjian. 1991. Transcription from a TATA-less promoter requires a multisubunit TFIID complex. Genes Dev 5:1935-45.

57.Quinlivan, E. B., E. A. Holley-Guthrie, M. Norris, D. Gutsch, S. L. Bachenheimer, and S. C. Kenney. 1993. Direct BRLF1 binding is required for cooperative BZLF1/BRLF1 activation of the Epstein-Barr virus early promoter, BMRF1. Nucleic Acids Res 21:1999-2007.

58.Ragoczy, T., and G. Miller. 2001. Autostimulation of the Epstein-Barr virus BRLF1 promoter is mediated through consensus Sp1 and Sp3 binding sites. J Virol 75:5240-51.

59.Robertson, E. S., B. Tomkinson, and E. Kieff. 1994. An Epstein-Barr virus with a 58-kilobase-pair deletion that includes BARF0 transforms B lymphocytes in vitro. J Virol 68:1449-58.

60.Saluja, D., M. F. Vassallo, and N. Tanese. 1998. Distinct subdomains of human TAFII130 are required for interactions with glutamine-rich transcriptional activators. Mol Cell Biol 18:5734-43.

61.Shimohata, M., T. Shimohata, S. Igarashi, S. Naruse, and S. Tsuji. 2005. Interference of CREB-dependent transcriptional activation by expanded polyglutamine stretches--augmentation of transcriptional activation as a potential therapeutic strategy for polyglutamine diseases. J Neurochem 93:654-63.

62.Smith, P. R., Y. Gao, L. Karran, M. D. Jones, D. Snudden, and B. E. Griffin. 1993. Complex nature of the major viral polyadenylated transcripts in Epstein-Barr virus-associated tumors. J Virol 67:3217-25.

63.Swaminathan, S., B. Tomkinson, and E. Kieff. 1991. Recombinant Epstein-Barr virus with small RNA (EBER) genes deleted transforms lymphocytes and replicates in vitro. Proc Natl Acad Sci U S A 88:1546-50.

64.Tanese, N., D. Saluja, M. F. Vassallo, J. L. Chen, and A. Admon. 1996. Molecular cloning and analysis of two subunits of the human TFIID complex: hTAFII130 and hTAFII100. Proc Natl Acad Sci U S A 93:13611-6.

65.Tjian, R., and T. Maniatis. 1994. Transcriptional activation: a complex puzzle with few easy pieces. Cell 77:5-8.

66.Tomkinson, B., and E. Kieff. 1992. Second-site homologous recombination in Epstein-Barr virus: insertion of type 1 EBNA 3 genes in place of type 2 has no effect on in vitro infection. J Virol 66:780-9.

67.Tomkinson, B., E. Robertson, and E. Kieff. 1993. Epstein-Barr virus nuclear proteins EBNA-3A and EBNA-3C are essential for B-lymphocyte growth transformation. J Virol 67:2014-25.

68.Weis, L., and D. Reinberg. 1992. Transcription by RNA polymerase II: initiator-directed formation of transcription-competent complexes. Faseb J 6:3300-9.

69.Wolf, H., H. zur Hausen, and V. Becker. 1973. EB viral genomes in epithelial nasopharyngeal carcinoma cells. Nat New Biol 244:245-7.

70.Wright, K. J., M. T. Marr, 2nd, and R. Tjian. 2006. TAF4 nucleates a core subcomplex of TFIID and mediates activated transcription from a TATA-less promoter. Proc Natl Acad Sci U S A 103:12347-52.

71.Zawel, L., and D. Reinberg. 1993. Initiation of transcription by RNA polymerase II: a multi-step process. Prog Nucleic Acid Res Mol Biol 44:67-108.

72.Zerby, D., C. J. Chen, E. Poon, D. Lee, R. Shiekhattar, and P. M. Lieberman. 1999. The amino-terminal C/H1 domain of CREB binding protein mediates zta transcriptional activation of latent Epstein-Barr virus. Mol Cell Biol 19:1617-26.