EB病毒是一種人類疱疹病毒，全世界有九成以上成人感染過，並造成終生的潛伏，隨後發現病毒的感染和許多癌症以及淋巴疾病有著高度的相關性。潛伏性膜蛋白1（LMP1）在EB病毒第一型及第二型潛伏期均有表現，是個具有多功能的蛋白，能造成囓齒動物的纖維母細胞轉型，並使得被EB病毒感染的B細胞不死，被認為是個致癌蛋白。LMP1的功能就像是個持續活化的腫瘤壞死因子受體，可以在不需配體的情況下活化許多訊息傳遞路徑，因而誘導許多基因的表現包括抗細胞凋亡及細胞素等。在許多不同的細胞LMP1的分佈是不全然相同的。為了確認它的位置，我們將LMP1與綠色螢光蛋白做融合表現，在三株上皮細胞觀察到它位於細胞膜上並有一大部分是聚集於細胞核週邊的部分。在次細胞層級，我們構築一系列能在特定胞器表現的紅色螢光蛋白作為標誌，這些胞器包括內質網、高基氏體以及細胞膜，我們發現LMP1在內質網、高基氏體以及細胞膜都有分佈。另一方面，我們利用兩套有效的siRNA篩選系統，成功的找到一個極為有效的siRNA分子，能夠非常專一的抑制LMP1的表現。這兩套系統一個是利用LMP1的cDNA與報告基因做融合的方式，另一系統是只將LMP1的標的序列與報告基因做融合表現。經由西方點墨、免疫螢光染色以及北方點墨的檢驗，我們確認有效的siRNA分子能專一的抑制LMP1蛋白質與mRNA的產生。進一步在穩定表現LMP1的細胞株裡，該有效的siRNA分子也能將LMP1的表現明顯降低。根據這些結果此有效的siRNA分子對於往後LMP1功能的研究將是一個有力的工具。

Epstein-Barr virus (EBV), a human herpesvirus that establishes a life-long persistent infection in more than 90% of adults worldwide, is highly associated with many tumors and lymphoid disease. Latent membrane protein 1 (LMP1), a versatile protein, not only involves in type II and type III EBV latency but also regards as an oncoprotein in rodent fibroblast transformation and EBV-induced B cell immortalization. LMP1 functions as a constitutively activated member of the tumor necrosis factor receptor (TNFR) superfamily and activates several signaling pathways in a ligand-independent manner, which induces various genes that encode anti-apoptotic proteins and cytokines. The distribution of LMP1 was shown to be heterogeneous among individual cells. To identify its cellular localization, we have expressed LMP1 fused with enhanced green fluorescence protein (EGFP), and in cell membrane and the most condensed in perinuclear compartments were found in three epithelial cell lines. Subcellular localizations of the LMP1 were determined with a series of organelle-specific markers for endoplasmic reticulum, Golgi apparatus and cell membrane. By construction, those organelle targeting sequences were reported with red fluorescence protein (DsRed2). We found that a proportion of LMP1 was co-localized with ER, Golgi apparatus and cell membrane. Besides, we had identified a potent siRNA molecule by using two efficient siRNA screening strategy. One is fused LMP1 cDNA with reporter genes, and the other is fused only siRNA target sequence with reporter genes. Using western blot, immunofluorescence staining and northern blot, the potent siRNA can inhibit LMP1 protein and mRNA production specifically. Furthermore, the potent siRNA reduced LMP1 expression in LMP1 transfected stable cell lines. These results suggested the potent siRNA molecular could be a power tool to study LMP1 functions.

Adriaenssens, E., Mougel, A., Goormachtigh, G., Loing, E., Fafeur, V., Auriault, C., Coll, J. (2004) A novel dominant-negative mutant form of Epstein-Barr virus latent membrane protein-1 (LMP1) selectively and differentially impairs LMP1 and TNF signaling pathways. Oncogene 23: 2681-2693.
Alberi, L., Sgado, P., and Simon, H.H. (2004) Engrailed genes are cell-autonomously required two prevent apoptosis in mesencephalic dopaminergic neurons. Development 131: 3229-3236.
Anderson, J., Banerjea, A., Planelles, V., and Akkina, R.(2003) Potent suppression of HIV type 1 infection by a short hairpin anti-CXCR4 siRNA. AIDS Res Hum Retroviruses 19: 699-706.
Angell, S.M., and Baulcombe, D.C. (1997) Consistent gene silencing in transgenic plants expressing a replicating potato virus X RNA. EMBO J. 16: 3675-3684.
Allday, M.J., and Crawford, D.H. (1988) Role of epithelium in EBV persistence and pathogenesis of B-cell tumours. Lancet 1: 855-857.
Baer, R., Bankier, A.T., Biggin, M.D., Deininger, P.L., Farrell, P.J., Gibson, T.J., Hatfull, G., Hudson, G.S., Satchwell, S.C., Seguin, C., and et al. (1984) DNA sequence and expression of the B95-8 Epstein-Barr virus genome. Nature 310: 207-211.
Bartel, D. P. (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116:281-297.
Bodary, S.C., and McLean, J.W. (1990) The integrin beta 1 subunit associates with the vitronectin receptor alpha v subunit to form a novel vitronectin receptor in a human embryonic kidney cell line. J Bio Chem 265: 5938-5941.
Braasch, D.A., Paroo, Z., Constantinescu, A., Ren, G., Oz, O.K., Mason, R.P., and et al. (2004) Biodistribution of phosphodiester and phosphorothioate SiRNA. Bioorg Med Chem Lett 14: 1139-1143.
Brennan, P., Floettmann, J.E., Mehl, A., Jones, M., and Rowe, M. (2001) Mechanism of action of a novel latent membrane protein-1 dominant negative. J Biol Chem 276: 1195-1203.
Brown, D.A., and London, E., (1998) Functions of lipid rafts in biological membranes. Annu Rev Cell Dev Biol 14: 111-136.
Brown, D.A., and London, E., (1998) Structure and Function of Sphingolipid- and Cholesterol-rich Membrane Rafts. J Biol Chem 275: 17221-17224
Brummelkamp, T.R., Bernards, R., and Agami, R. (2002) A System for Stable Expression of Short Interfering RNAs in Mammalian Cells. Science 296: 550-553.
Burke, A.P., Yen, T.S., Shekitka, K.M. and Sobin, L.H. (1990) Lymphoepithelial carcinoma of the stomach with Epstein-Barr virus demonstrated by polymerase chain reaction. Mod. Pathol. 3: 377-380.
Burkitt, D. (1958) A carcoma involving the jaw in African children. Brit. J. Surg. 16: 218-223.
Busch, L.K., Bishop, G.A. (1999) The EBV transforming protein, latent membrane protein 1, mimics and cooperates with CD40 signaling in B lymphocytes. J Immunol 162: 2555-2561.
Cahir-McFarland, E.D., Davidson, D.M., Schauer, S.L., Duong, J., and Kieff, E. (2000) NF-κB inhibition causes spontaneous apoptosis in Epstein-Barr virus-transformed lymphoblastoid cell lines. Proc Natl Acad Sci USA 97: 6055-6060.
Chatterjee, S., Smith, E.R., Hanada, K., Stevens, V.L., and Mayor, S. (2001) GPI anchoring leads to sphingolipid-dependent retention of endocytosed proteins in the recycling endosomal compartment. EMBO J 20: 1583-1592.
Chen CJ, Chang CL, Liaw YP, Shia CF, Huang KL, You SL, and et al. (1997) Atlas of Cancer Mortality in Taiwan, 1982–1991.
Chen, C.J., You, S.L., Lin, L.H., Hsu, W.L., and Yang, Y.W. (2002) Cancer Epidemiology and Control in Taiwan: a Brief Review. Jpn J Clin Oncol. 32: S66-81.
Chen, H., Lee, J.M., Zong, Y., Borowitz, M., Ng, M.H., Ambinder, R.F., Hayward, S.D. (2001) Linkage between STAT regulation and Epstein-Barr virus gene expressionin tumors. J Virol 75: 2929-2937.
Chi, J.T., Chang, H.Y., Wang, N.N., Chang, D.S., Dunphy, N., Brown, P.O. (2003) Genomewide view of gene silencing by small interfering RNAs. Proc. Nat. Acad. Sci. USA 100: 6343-6346.
Clausse, B., Fizazi, K., Walczak, V., Tetaud, C., Wiels, J., Tursz, T., and Busson, P. (1997) High concentration of the EBV latent membrane protein 1 in glycosphingolipid-rich complexes from both epithelial and lymphoid cells. Virolgy 228: 285-293.
Coffin, W.F. 3rd., Erickson, K.D., Hoedt-Miller, M., and Martin, J.M. (2001) The cytoplasmic amino-terminus of the Latent Membrane Protein-1 of Epstein- Barr Virus: Relationship between transmembrane orientation and effector functions of the carboxy-terminus and transmembrane domain. Oncogene 20: 5313-5330.
Cogoni, C.M., and Macino, G. (1997) Isolation of quelling-defective (QDE) mutants impaired in posttranscriptional transgene-induced gene silencing in Neurospora crassa. Proc. Nat. Acad. Sci. USA 94: 10233-10238.
Dambaugh, T., Beisel, C., Hummel, M., King, W., Fennewald, S., Cheung, A., Hellen, M., Raab-Traub, N., and Kieff, E. (1980) Epstein-Barr virus (B95-8) DNA VII: molecular cloning and detailed mapping. Proc Natl Acad Sci USA. 77: 2999-3003.
Dawson, C.W., Rickinson, A.B., and Young, L.S. (1990) Epstein–Barr virus latent membrane protein inhibits human epithelial cell differentiation. Nature 344: 777-780.
Dernburg, A.F., Zalevsky, J., Colaiacovo, M.P., and Villeneuve, A.M. (2000) Transgene-mediated cosuppression in the C. elegans germ line. Genes Dev. 14: 1578-1583 .
Devergne, O., Hatzivassiliou, E., Izumi, K.M., Kaye, K.M., Kleijner, M., Kieff, E., and Mosialos, G. (1996) TRAF1, TRAF2 and TRAF3 effect NF-κB activation by an Epstein-Barr Virus LMP1 domain important for B lymphocyte transformation. Mol Cell Biol. 16: 7098-7107.
D’Souza, B., Rowe, M., and Walls, D. (2000) The bfl-1 gene is transcriptionally upregulated by the Epstein–Barr virus oncoprotein LMP1, and its expression promotes the survival of a Burkitt’s lymphoma cell line. J Virol 74: 6652–6658.
Elbashir, S.M., Harborth, J., Lendeckel, W., Yalcin, A., Weber, K., ant Tuschel, T. (2001) Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 411: 494-498.
Eliopoulos, A.G., Gallagher, N.J., Blake, S.M.S., Dawson, C.W., and Young. L.S. (1999) Activation of the p38 MAPK pathway by Epstein–Barr virus encoded latent membrane protein 1(LMP1) co-regulates interleukin-6 and interleukin-8 production. J Biol Chem 274: 16085–16096.
Eliopoulos, A.G., and Young. L.S. (1998) Activation of the cJun N-terminal kinase (JNK) pathway by the Epstein-Barr virus-encoded latent membrane protein 1 (LMP1). Oncogene 16: 1731-1742.
Epstein, M.A., and Barr, Y.M. (1964) Cultivation in vitro of human lymphoblast from Burkitt’s malignant lymphoma. Lancent. 1: 252-253.
Epstein, M.A., Achong, B.G., and Barr, Y.M. (1964) Viral particles in cultured lymphoblasts from Burkitt’s lymphoma. Lancet 1: 702-703.
Fahraeus, R., Chen, W., Trivedi, P., Klein, G., Obrink, B. (1992) Decreased expression of E-cadherin and increased invasive capacity in EBV-LMP1-transfected human epithelial and murine adenocarcinoma cells. Int J Cancer 52: 834-838.
Fahraeus, R., Fu, H.L., Emberg, I., Finke, J., Rowe, M., Klein, G., Falk, K., Nilsson, E., Yadav, M., Busson, P., and et al. (1988) Expression of Epstein-Barr virus- encoded protein in nasopharyngeal carcinoma. Int J Cancer 42: 329-338.
Fahraeus, R., Rymo, L., Rhim, J.S., Klein, G. (1990) Morphological transformation of human keratinocytes expressing the LMP gene of Epstein-Barr virus. Nature 345: 447-449.
Fennewald, S.V., van Santen, V., and Kieff, E. (1984) Nucleotide sequence of an mRNA transcribed in latent growth-transforming virus infection indicates that it may encode a membrance protein. J Virol 51: 411-419.
Fire, A., and et al. (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391: 806-811.
Flanagan, J., Middeldorp, J., and Sculley, T. (2003) Localization of the Epstein-Barr virus protein LMP 1 to exosomes. J Gen Virol 84: 1871-1879
Fliegel, L., Burns, K., MacLennan, D.H., Reithmeier, R.A., and Michalak, M. (1989) Molecular cloning of the high affinity calcium-binding protein (calreticulin) of skeletal muscle sarcoplasmic reticulum. J Biol Chem 264: 21522 -21528.
Floettmann, J.E., Ward, K., Rickinson, A.B., and Rowe,M. (1996) Cytostatic effect of Epstein-Barr virus latent membrane protein-1 analyzed using tetracycline-regulated expression in B cell lines. Virology 223: 29-40.
Ge, Q., McManus, T., Nguyen, T., Shen, C.H., Sharp, P.A., Eisen, H.N., and et al.(2003) RNA interference of influenza virus production by directly targeting mRNA for degradation and indirectly inhibiting all viral RNA transcription. Proc Natl Acad Sci USA 100: 2718-23.
Gertrude Henle and Werner Henle (1966) Immunofluorescence in Cells Derived from Burkitt's Lymphoma. J Bacteriol. 91: 1248-1256.
Gires, O., Kohhuber. F., Kilger. E., Baumann. M., Kieser. A., Kaiser, C., Zeidler, R.,Scheffer, B., Ueffing, M., and Hammerschmidt, W. (1999) Latent membrane protein 1 of Epstein-Barr virus interacts with JAK3 and activates STAT proteins. EMBO 18: 3064-3073.
Gires, O., Zimber-Strobl, U., Gonnella, R., Ueffing, M., Marschall, G., Zeidler, R., Pich, D., and Hammerschmidt, W. (1997) Latent membrane protein 1 of Epstein-Barr virus mimics a constitutively active receptor molecule. EMBO J 16: 6131-6140.
Glaser, R., Ogino, T., Zimmerman, J., Jr., and Rapp, F. (1973) Thymidine kinase activity in Burkitt lymphoblastoid somatic cell hybrids after induction of the EB virus. Proc Soc Exp Biol Med 142: 1059-62.
Glaser, R., Zhang, H.Y., Yao, K., Zhu, H.C., Wang, F.X., Li, G.Y., Wen, D.S., and Li Y.P. (1989) Two epithelial tumor cell lines (HNE-1 and HONE-1) latently infected with Epstein-Barr virus that were derived from nasopharyngeal carcinomas. Proc Natl Acad Sci USA 86: 9524-9528.
Gleave, M.E., Monia, B.P. (2005) Antisense therapy for cancer. Nat Rev Cancer 5: 468-478.
Graham, F.L., Smiley, I., Russell, W.C., and Nairn, R. (1977) Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol 36: 59-74.
Gratama, J.W., Zutter, M.M., Minarovits, J., Oosterveer, M.A., Thomas, E.D., Klein, G., and Ernberg, I. (1991) Expression of Epstein-Barr virus-encoded growth transformation-associated proteins in lymphoproliferations of bone-marrow transplant recipients. Int J Cancer 47: 188-192.
Greenspan, J.S., Greenspan, D., Lennette, E.T., Abrams, D.I., Conant, M.A., Petersen, V., and Freese, U.K. (1985) Replication of Epstein-Barr virus within the epithelial cells of oral “hairy“ leukoplakia, an AIDS-associated lesion. N. Engl. J Med. 313: 1564-1571.
Guo, S., and Kemphues, K.J. (1995) par-1, a gene required for establishing polarity in C. elegans embryos, encodes a putative Ser/Thr kinase that is asymmetrically distributed. Cell 81: 611-620.
Hammerschmidt, W., Sugden. B., Baichwal, V.R. (1989) The transforming domain alone of the latent membrane protein of Epstein-Barr virus is toxic to cells when expressed at high levels. J Virol 63: 2469-2475.
Hannon, G.J. (2002) RNA interference. Nature 418: 244–51.
He, Z., Xin, B., Yang, X., Chan, C., and Cao, L. (2000) Nuclear factor-kappaB activation is involved in LMP1-mediated transformation and tumorigenesis of rat-1 fibroblasts. Cancer Res 60: 1845-1848.
Heino, S., Lusa, S., Somerharju, P., Ehnholm, C., Olkkonen, V.M., and Ikonen, E. (2000) Dissecting the role of the Golgi complex and lipid rafts in biosynthetic transport of cholesterol to the cell surface. Proc Natl Acad Sci USA 97: 8375-8380.
Henderson, S., Rowe, M., Gregory, C., Croom-Carter, D., Wang, F., Longnecker, R., Kieff, E., and Rickinson, A.B. (1991) Induction of bcl-2 expression by the Epstein-Barr-Virus Latent Membrane Protein-1 protects infected B-cells from programmed cell death. Cell 65: 1107-1115.
Henle, G., and Henle, W. (1966) Immunofluorescence in cells derived from Burkitt’s lymphoma. J. Bacteriol. 91: 1248-1256.
Henle, G., Henle, W., and Diehl, V. (1968) Relation of Burkitt’s tumor-associated herpes-type virus to infectious mononucleosis. Proc Natl Acad Sci USA 59: 94-101.
Hennessy, K., Fennewald, S., Hummel, M., Cole, T., and Kieff, E. (1984) A membrane protein encoded by Epstein-Barr virus in latent growth transforming infection. Proc Natl Acad Sci USA 81: 7202-7211.
Henriquez, N.V., Floettmann, E., Salmon, M., Rowe, M., Rickinson, A.B. (1999) Differential responsets to CD40 ligation among Burkitt lymphoma lines that are uniformly responsive to Epstein-Barr virus latent membrane protein 1. J Immunol 162: 3298-3307.
Horikawa, T.,Yoshizaki, T.,Sheen, T.S., Lee, S.Y., and Furukawa, M.(2000) Association of latent membrane protein 1 and matrix metalloproteinase 9 with metastasis in nasopharyngeal carcinoma. Cancer 89: 715-723.
Horikawa, T., Sheen, T.S., Takeshita, H., Sato, H., Furukawa, M., and Yoshizaki, T. (2001) Induction of c-Met protooncogene by Epstein-Barr virus latent membrane protein-1 and the correlation with cervical lymph node metastasis of nasopharyngeal carcinoma. Am J Pathol 159:33.
Howard, K. (2003) Unlocking the money-making potential of RNAi. Nat Biotechnol 21: 1441-1446.
Huang, F., Khvorova, A., Marshall, W., and Sorkin, A. (2004) Analysis of clathrin-mediated endocytosis of epidermal groth factor receptor by RNA interference. J Biol Chem 279: 16657-16661.
Huen, D.S., Henderson, S.A., Croom-Carter, D., and Rowe, M. (1995) The Epstein- Barr Virus latent membrane protein 1 (LMP1) mediates activation of NF-κB and cell surface phenotype via two effector regions in its carboxy-terminal cytoplasmic domain. Oncogene 10: 549-560.
Ikonen, E. (2001) Roles of lipid rafts in membrane transport. Curr Opin Cell Biol 13: 470–477.
Ingelbrecht, I., Van Houdt, H., Van Montagu, M., and Depicker, A. (1994) Post- transcriptional silencing of reporter transgenes in tobacco correlates with DNA methylation. Proc. Natl Acad. Sci. USA 91: 10502-10506.
Izumi, K.M., Kaye, K.M., and Kieff, E.D. (1994) Epstein-Barr virus recombinant molecular genetic analysis of the LMP1 amino-terminal cytoplasmic domain reveals a probable structural role, with no component essential for primary B-lymphocyte growth transformation. J Virol 68: 4369-4376.
Izumi, K.M., McFarland, E.C., Ting, A.T., Riley, E., Seed, B., and Kieff, E.D. (1999) The Epstein-Barr virus oncoprotein Latent Membrane protein 1 engages the tumour necrosis factor receptor-associated proteins TRADD and receptor interacting protein (RIP) but does not induce apoptosis or require RIP for NF-κB activation. Mol Cell Biol. 19: 5759-5767.
Jeffrey, I., and Cohen, M.D. (2000) Epstein–Barr virus infection. N. Engl. J. Med. 343: 481-492.
Jones, J.F., Shurin, S., Abranowaky, C., Tubbs, R.R., Sciotto, C.G., Wahl, R., Sands, J., Gottman, D., Ktaz, B.Z., and Sklar, J. (1988) T-cell lymphoma containing Epstein-Barr viral DNA in patients with chronic Epstein-Barr virus infections. N. Engl. J Med. 318: 733-741.
Kartasheva, N.N., Contente, A., Lenz-Stoppler, C., Roth, J., and Dobbelstein, M. (2002) P53 induces the expression of its antagonist p73 delta N, establishing an autoregulatory feedback loop. Oncogene 21: 4715-4721.
Kaye, K.M., Izumi, K.M., and Kieff, E. (1993) Epstein-Barr virus latent membrane protein 1 is essential for B-lymphocyte growth transformation. Proc Natl Acad Sci USA 90: 9150-9154.
Kaye, K.M., Izumi, K.M., Li, H., Johannsen, E., Davidson, D., Longnecker, R., and Kieff, E. (1999) An Epstein-Barr virus that expresses only the first 231 LMP1 amino acids efficiently initiates primary B-lymphocyte growth transformation. J. Virol. 73: 10525-10530.
Kaye, K.M., Izumi, K.M., Mosialos, G., and Kieff, E. (1995) The Epstein-Barr virus LMP1 cytoplasmic carboxy terminus is essential for B lymphocyte transformation; fibroblast cocultivation complements a critical function within the terminal 155 residues. J. Virol. 69: 675-683.
Kaykas, A., and Sugden, B. (2000) The amino-terminus and membrane-spanning domains of LMP-1 inhibit cell proliferation. Oncogene 19: 1400-1410.
Kaykas, A., Worringer, K., and Sugden, B. (2001) CD40 and LMP-1 both signal from lipid rafts but LMP-1 assembles a distinct, more efficient signaling complex. EMBO J 20: 2641-2654.
Kenney, J.L., Guinness, M.E., Curiel, T., and Lacy, J. (1998) Antisense to the Epstein-Barr virus (EBV)-encoded latent membrane protein 1 (LMP-1) suppresses LMP-1 and bcl-2 expression and promotes apoptosis in EBV-immortalized B cell. Blood 92 1721-1727.
Kikuta, H., Taguchi, Y., Tomizawa, K., Kijima, K., Kawamura, N., Ihizaka, A., Sakiya, Y., Matsumoto, S., Imai, S., Kinoshita, T., and et al. (1988) Epstein-Barr virus genome-positive T lymphocytes in a boy with chronic active EBV infection associated with Kawasaki-like disease. Nature 333: 455-457.
Kilger, E., Kieser, A., Baumann, M., and Hammerschmidt, W. (1998) Epstein-Barr virus-mediated B-cell proliferation is dependent upon latent membrane protein 1, which simulates an activated CD40 receptor. EMBO J 17: 1700-1709.
Komano, J., Sugiura, M., and Takada, K. (1998) Epstein-Barr virus contributes to the malignant phenotype and to apoptosis resistence in Burkitt’s lymphoma cell line Akata. J Virol 72: 9150-9156.
Lam, N., and Sugden, B. (2002) CD40 and its viral mimic, LMP1: similar means to different ends. Cell Signal 15: 9-16.
Lam, N., and Sugden, B. (2003) LMP1, a viral relative of the TNF receptor family, signals principally from intracellular compartments. EMBO 22: 3027-3038.
Laherty, C.D., Hu ,H.M., Opipari, A.W., Wang, F., and Dixit, V.M. (1992) The Epstein–Barr Virus LMP-1 product induces A20 zinc finger protein expression by activating nuclear factor-B. J Biol Chem 267: 24157-24160.
Laszlo, L., Tuckwell, J., Self, T., Lowe, J., Landon, M., Smith, S., Hawthorne, J. N., and Mayer, R. J. (1991) The latent membrane protein-1 in Epstein–Barr virus-transformed lymphoblastoid cells is found with ubiquitin-protein conjugates and heat-shock protein 70 in lysosomes oriented around the microtubule organizing centre. J Pathol 164: 203-214.
Lee, M.T., Coburn, G.A., McClure, M.O., and Cullen, B.R. (2003) Inhibition of human immunodeficiency virus type-1 replication in primary macrophages by using Tat- or CCR5-specific small interfering RNAs expressed from a lentivirus vector. J Virol 77: 11964-11972.
Lee, R.C., Feinbaum, R.L., and Ambros, V. (1993) The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. Cell 75: 843-54.
Li, Q., Spriggs, M.K., Kovats, S., Turk, S.M., Comeau, M.R., Nepom, B., and Hutt-Fletcher, L.M. (1997) Epstein-Barr virus uses HLA class II as a cofactor for infection of B lymphocytes. J. Virol. 71: 4657-4662.
Li, X.P., Li, G., Peng, Y., Kung, H., and Lin, M.C. (2004) Suppression of Epstein-Barr virus-encoded latent membrane protein-1 by RNA interference inhibits the metastatic potential of nasopharyngeal carcinoma cells. Biochem Biophys Res Commun 315: 212-218.
Liebowitz, D., and Kieff, E. (1989) Epstein-Barr virus latent membrane protein: induction of B-cell activation antigens and membrane patch formation does not require vimetnin. J. Virol. 63: 4051-4054.
Liebowitz, D., Kopan, R., Fuchs, E., Sample, J., and Kieff, E. (1987) An Epstein-Barr virus transforming protein associates with vimentin in lymphocyts. Mol. Cel. Biol. 7: 2299-2308.
Liebowitz, D., Wang, D., and Kieff, E. (1986) Orientation and patching of the latent membrane protein encoded by Epstein-Barr virus. J Virol 58:233-237.
Linde, A. (1996) Diagnosis of Epstein-Barr virus-related diseases. Scand. J. Infect. Dis. 28: 83-88.
Liu, Y., Wang, X., Lo, A.K., Wong, Y.C., Cheung, A.L., Tsao, S.W. (2002) Latent membrane protein-1 of Epstein-Barr virus inhibits cell growth and induces sensitivity to cisplatin in nasopharyngeal carcinoma cells. J Med Virol 66: 63-69.
Llopis, J., Michael McCaffery,J., Miyawaki, A., Farquhar, M.G., and Tsien, R.Y. (1998) Measurement of cytosolic, mitochondrial, and Golgi pH in single living cells with green fluorescent proteins. PNAS 95: 6803-6808.
Luka, J., Kallin, B., and Klein, G. (1979) Induction of the Epstein –Barr virus (EBV) cycle in latently infected cells by n-butyrate. Virology 193: 228-231.
Mann, K.P., Staunton, D., and Thorley-Lawson, D.A. (1985) An Epstein-Barr virus-encoded protein found in plasma membranes of transformed cells. J Virol 55: 710-720.
Mann, K.P., and Thorley-Lawson, D. (1987) Posttranslational processing of the Epstein-Barr virus-encoded p63/LMP protein. J. Virol. 61: 2100-2108.
Mattia, E., Chichiarelli, S., Hickish, T., Gaeta, A., Mancini, C., Cunningham, D., and van Renswoude, J. (1997) Inhibition of in vitro proliferation of Epstein-Barr Virus infected B cells by an antisense oligodeoxynucleotide targeted against EBV latent membrane protein LMP1. Oncogene 15: 489-493.
Macpherson, I. and Stoker, M. (1962) Polyoma transformation of hamster cell clones--an investigation of genetic factors affecting cell competence. Virology 16: 147-151.
Miller, G., and Lipman, M. (1973) Release of infectious Epstein-Barr virus by transformed marmoset leukocytes. Proc Natl Acad Sci U S A 70: 190-194.
Min, K.W., Holmquist, S., Peiper, S.C., and O’Leary, T.L. (1991) Poorly differentiated adenocarcinoma with lymphoid stroma （lymphoepithelioma-like carcinoma）of the stomach. Report of three cases with Epstein-Barr virus genomes demonstrated by the polymerase chain reaction. Am. J. Clin. Pathol. 96: 219-227.
Mitchell, T., and Sudgen, B. (1995) Stimulation of NF-kB-mediated transcription by mutant derivatives of the latent membrane protein of Epstein-Barr virus. J. Virol. 69: 2968-2976.
Mittal, V. (2004) Improving the efficiency of RNA interference in mammals. Nature Review Genetics 5: 355-365.
Miyagishi, M., and Taira, K. (2002) U6-promoter driven siRNA with four uridine 3' overhangs efficiently surpass targeted gene expression in mammalian cells. Nature Biotechnology 20: 497-500.
Mosialos, G., Birkenbach, M., Yalamanchili, R., VanArsdale, T., Ware, C., and Kieff, E. (1995) The Epstein-Barr virus transforming protein LMP1engages signaling proteins for the tumor necrosis factor receptor family. Cell 80: 389-399.
Murray, R. J., Kurilla, M.G., Brooks, J. M., Thomas, W.A., Rowe, M., Kieff, E., and et al. (1992) Identification of target antigens for the human cytotoxic T-cell response to EBV: implications for the immune control of EBV-positive malignancies. J. Exp. Med. 176: 157-168.
Nakagomi, H., Dolcetti, R., Bejarano, M.T., Pisa, P., Kiessling, R., Masucci, M.G. (1994) The Epstein-Barr virus latent membrane protein-1 (LMP1) induces interleukin-10 production in Burkitt lymphoma lines. Int J Cancer 58: 240-244.
Napoli, C., Lemieux, C., and Jorgensen, R. (1990) lntroduction of a Chimeric Chalcone Synthase Gene into Petunia Results in Reversible Co-Suppression of Homologous Genes in trans. Plant Cell 2: 279-289.
Nemerow, G.R., Moore, M.D., and Cooper, N.R. (1990) Structure and function of the B-lymphocyte Epstein-Barr virus/C3d receptor. Adv. Cancer Res. 54: 273-300.
Noguchi, T., Ikeda, K., Yamamoto, K., Yoshida, I., Ashiba, A., and et al. (2001) Antisense oligodeoxynucleotides to latent membrane protein 1 induce growth inhibition, apoptosis and Bcl-2 suppression in Epstein-Barr virus (EBV)-transformed B-lymphoblastoid cells, but not in EBV-positive natural killer cell lymphoma cells. Br J Haematol 114: 84-92.
Novina, C.D., Murray, M.F., Dykxhoorn, D.M., Beresford, P.J., Riess, J., Lee, S.K., Collman, R.G., Lieberman, J., Shankar, P., and Sharp, P.A. (2002) siRNA-directed inhibition of HIV-1 infection. Nat. Med. 8: 681-686.
Niedobitek, G., Agathanggelou, A., Herbst, H., Whitehead, L., Wright, D.H., and Young, L.S. (1997) Epstein-Barr virus (EBV) infection in infectious mononucleosis: virus latency, replication and phenotype of EBV-infected cells. J Pathol 182: 151-9.
Pallesen, G.,Hamilton-Dutolt, S.J., Rowe, M., and Young, I.S. (1991) Epstein-Barr virus latent genes in tumour cells of Hodgkin’s diseas. Lancet 337: 320-322.
Pasquinelli, A.E., and Ruvkun, G. (2002) Control of developmental timing by micromas and their targets. Annu Rev Cell Dev Biol 18: 171-173.
Pathmanathan, R. , Prasad, U., Sadler, R., Flynn, K., and Raab-Traub, N. (1995) Clonal Proliferations of Cells Infected with Epstein–Barr Virus in Preinvasive Lesions related to Nasopharyngeal Carcinoma. N Engl J Med 333: 693-698.
Peng, M., and Lundgren, E. (1992) Transient expression of the Epstein–Barr virus LMP1 gene in human primary B cells induces cellular activation and DNA synthesis. Oncogene 7: 1775–1782.
Pfeffer, S., Zavolan, M., Grasser, F.A., Chien, M., Russo, J.J., Ju, J., John, B., Enright, A.J., Marks, D., Sander, C., and Tuschl, T. (2004) Identification of virus-encoded microRNAs. Science 304: 734–736.
Reynolds, A., Leake, D., Boese, Q., Scaringe, S., Marshall, W.S., and Khvorova, A. (2004) Rational siRNA design for RNA interference. Nat Biotechnol 22: 326-330.
Rossi, J.J. et al. (2005) Synthetic dsRNA Dicer substrates enhance RNAi potency and efficacy. Nat Biotechnol 23, in the press.
Roth, J., Claudia König, C., Wienzek, S., Weigel, S., Ristea, S., and Dobbelstein, M. (1998) Inactivation of p53 but Not p73 by Adenovirus Type 5 E1B 55-Kilodalton and E4 34-Kilodalton Oncoproteins. J Virol 72: 8510-8516.
Rowe, M., Rowe, D.T., Gregory, C.D., Young, L.S., Farrell, P.J., Rupani, H., and Rickinson, A.B. (1987) Differences in B cell growth phenotype reflect novel patterns of Epstein-Barr virus latent gene expression in Burkitt’s lymphoma cells. EMBO J. 6: 2743-51.
Sandberg, M., Hammerschmidt, W., and Sugden, B. (1997) Characterization of LMP-1’s association with TRAF1, TRAF2, and TRAF3. J Virol 71: 4649-4656.
Schultheiss, U., Puschner, S., Kremmer, E., W.Mak, T., Engelmann, H., Hammerschmidt, W., Kieser, A. (2001) TRAF6 is a critical mediator of signal transduction by the viral oncogene latent membrane protein 1. EMBO 20: 5678-5691.
Schwarz, D.S., Hutvagner, G., Haley, B., and Zamore, P.D. (2002) Evidence that siRNAs function as guides, not primers, in the Drosophila and human RNAi pathways. Mol Cell 10: 537-548.
Semizarov, D., Kroeger, P., and Fesik, S. (2004) SiRNA-mediated gene silencing: a global genome view. Nucleic Acids Res 32: 3836-3845.
Shu, X., Wu, W., Mosteller, R.D., and Broek, D. (2002) Sphingosine kinase mediates vascular endothelial growth factor- induced activation of ras and mitogen-activated protein kinases. Mol Cell Biol 22: 7758-7768.
Sheu, L.F., Chen, A.,Wei, Y.H., Ho, K.C., Cheng, J.Y., Meng, C.L., Lee, W.H. (1998) Epstein-Barr virus LMP1modulates the malignant potential of gastric carcinoma cells involving apoptosis. Am J Pathol 152: 63-74.
Siolas, D., Lerner, C., Burchard, J., Ge, W., Linsley, P.S., Paddison, P.J., Hannon, G.H., and Cleary, M.A. (2004) Synthetic shRNAs as potent RNAi triggers. Nature Biotechnolog 23: 227-231.
Sixbey, J.W., Vesterinen, E.H., Nedrud, J.G., Raab-Traub, N., Walton, L.A., and Pagano, J.S. (1983) Replication of Epstein-Barr virus in human epithelial cells infected in vitro. Nature 306: 480-483.
Skene, J.H., and Virag, I. (1989) Posttranslational membrane attachment and dynamic fatty acylation of a neuronal growth cone protein, GAP-43. J Cell Biol 108: 613-624.
Speck, P., Hann, K.M., Longnecker, R. (2000) Epstein-Barr virus Entry into Cells. Virology 277: 1-5.
Stark, G.R., Kerr, I.M., Williams, B.R., Silverman, R.H., and Schreiber, R.D. (1998) How cells respond to interferons. Annu Rev Biochem 67: 227-264.
Sun, L., Deng, L., Ea, C.K., Sia, Z.P., and Chen, Z.J. (2004) The TRAF6 ubiquitin ligase and TAK1 kinase mediate IKK activation by BCL10 and MALT1 in T lymphocytes. Mol Cell 14: 289-301.
Tabara, H., Sarkissian, M., Kelly, W.G., Fleenor, J., Grishok, A., Timmons, L., Fire, A., and Mello, C.C. (1999) The rde-1 gene, RNA interference, and transposon silencing in C. elegans. Cell 99: 123-132.
Thomas, J.A., Hotchin, N.A., Allday, M.J., Amlot, P., Rose, M., Yacoub, M., and Crawford, D.H. (1990) Immunohistology of EBV-associated antigens in B cell disorders from immunocompromised individuals. Transplantation 49: 944-953.
Tovey, M.G., Lenoir, G., and Begon-Lours, J. (1978) Activation of latent Epstein- Barr virus by antibody to human IgM. Nature 276: 270-272.
Tugizov, S.M., Berline, J.W., Palefsky, J.M. (2003) Epstein-Barr virus infection of polarized tongue and nasopharyngeal epithelial cells. Nature medicine 9: 307-314.
Vickers, T.A., Koo, S., Bennett, C.F., Crooke, S.T., Dean, N.M., and Baker, B.F. (2003) Efficient reduction of target RNAs by small interfering RNA and Rnase H-dependent antisense agents: A comparative analysis. J Biol Chem 278: 7108-7118.
Yoshizaki, T. (2002) Promotion of metastasis in nasopharyngeal carcinoma by Epstein-Barr virus latent membrane protein-1. Histol Histopathol 17: 845-850.
Wang, D., Liebowitz, D., Kieff, E. (1985) An EBV membrane protein expressed in immortalized lymphocytes transforms established rodent cells. Cell 43: 831-840.
Wang, S., Rowe, M., and Lundgren, E. (1996) Expression of the Epstein–Barr virus transforming protein LMP1 causes a rapid and transient stimulation of the Bcl-2 homologue Mcl-1 levels in B cell lines. Cancer Res. 56: 4610-4613.
Weiss, L.M., Movahed, L.A., Warnke, R.A., and Sklar, J. (1989) Detection of Epstein-Barr viral genomes in Reed-Stemberg cells of Hodgkin’s disease. N. Engl. J Med. 320: 502-506.
William, F. Coffin III, Kimberly, D. Erickson, Marloes Hoedt-Miller and Jennifer, M. Martin (2001) The cytoplasmic amino-terminus of the Latent Membrane Protein-1 of Epstein-Barr Virus: relationship between transmembrane orientation and effector functions of the carboxy-terminus and transmembrane domain. Oncogene 20: 5313-5330.
Wolf, J.B., and Seibel, R. (1984) Benign and malignant disease caused by EBV. J Invest. Dermatol. 83: 88s-95s.
Wu, M.T., Wu R.H., Hung, C.F., Cheng, T.L., Tsai, W.H., Chang, W.T. (2004) Simple and efficient DNA vactor based RNAi systems in mammalian cells. Biochem Biophys Res Commun 330: 53-59.
Yang, X., He, Z., Xin, B., and Cao, L. (2000) LMP1 of Epstein–Barr virus suppresses cellular senescence associated with the inhibition of p16 (INK4a) expression. Oncogene 19: 2002-2013.
Yoshizaki, T., Sato, H., Furukawa, M., and Pagano, J.S. (1998) The expression of matrix metalloproteinase 9 is enhanced by Epstein–Barr virus latent membrane protein 1. Proc Natl Acad Sci USA 95: 3621-3626.
Young, L., Alfieri, C.,Hennessy, K.,Evans, H., O’Hara, C., Anderson, K.C., Ritz. J., Shapiro, R.S., Rickinson, A., Kieff, E., and Cohen, J.I. (1989) Expression of EBV transformation-associated genes in tissues of patients with EBV lymphoproliferative disease. N Engl J Med 321: 1080-1085.
Yu, J.Y., DeRuiter, S.L., and Turner, D.L. (2002) RNA interference by expression of short interfering RNAs and hairpin RNAs in mammalian cells. Proceedings of the National Academy of Science of the USA 99: 6047-6052.
Zheng, L., Liu, J., Batalov, S.,Zhou, D., Orth, A., Ding. S., and Schultz, P.G. (2004) An approach to genomewide screens of expressed small interfering RNAs in mammalian cells. Proc Natl Acad Sci USA 101: 135-140.
Ziegler, J.L., Drew, W.L., Miner, R.C., Mintz, L., Rosenbaum, E., Gershow, J., Lennette, E.T., Greenspan, J., Shillitoe, E., Beckstead, J., Casavant, C., and Yamamoto, K. (1982) Outbreak of Burkitt;’s-like lymphoma in homosexual men. Lancet 2: 631-633.
Zur Hausen, H., Schulte-Holthausen, H., Klein, G., Henle, P., Clifford, P., Santesson, L. (1970) EBV DNA in biopsies of Burkitt tumours and anaplastic carcinomas of the nasopharynx. Nature 228: 1056-1058.