Molecular mechanism of Epstein-Barr virus latent infection

EB病毒潜伏感染的分子机制

• 批准号: 14570274
• 负责人: HARADA Shizuko
• 金额: $ 2.56万
• 依托单位: National Institute of Infectious Disease
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570274/
• 关键词: EBV; virus; latent infection; nuclear protein; transactivation; EBNA; ドミナントネガティブ

Epstein-Barr virus (EBV) is a ubiquitous human herpesvirus that latently infects B-lymphocytes and causes their continuous proliferation into lymphoblastoid cell lines (LCL). EBV nuclear protein EBNA-2 and EBNA-LP are the first viral proteins expressed in lymphocytes. They cooperatively up-regulate EBV and cellular gene transcription, and are required for lymphocyte outgrowth into LCL. We have now found that EBNA-LP readily associated with EBNA2 or with the EBNA2 C-terminal activation domain (E2AD) when both components were expressed in bacteria. In lymphoblasts, EBNA-LP and EBNA2 did not stably associate. However, EBNA-LP deleted for only 10 C-terminal amino acids (aa) stably associated with EBNA2 in lympohoblasts. EBNA-LP 31 aa (dW2Y1) with 24 C-or N-terminal aa was a specific and efficient affinity matrix for EBNA2 or EBNA-LP. These biochemical interactions between EBNA-LP and EBNA2 enable coordinated transcriptional regulation of cell and viral gene expression in lymphoblasts only when the interaction is unstable ; deletion of the EBNA-LP C-terminal 10aa stabilizes association with EBNA2 and prevents co-activation. Since EBNA-LPd10 dominantly inhibited EBNA-LP co-activation with EBNA2, EBNA-LP d10 over-expression in LCLs may be useful in assessing the role of EBNA-LP co-activation in LCL growth or survival.

eb病毒（Epstein-Barr virus， EBV）是一种普遍存在的人类疱疹病毒，可潜伏感染b淋巴细胞并使其持续增殖为淋巴母细胞样细胞系（LCL）。EBV核蛋白EBNA-2和EBNA-LP是在淋巴细胞中首先表达的病毒蛋白。它们协同上调EBV和细胞基因转录，是淋巴细胞生长为LCL所必需的。我们现在已经发现，当EBNA-LP和EBNA2 c -末端激活域（E2AD）在细菌中表达时，它们很容易与EBNA2或EBNA2 c -末端激活域（E2AD）结合。在淋巴细胞中，ena - lp和EBNA2没有稳定的关联。然而，在淋巴细胞中，ena - lp只删除了10个与EBNA2稳定相关的c端氨基酸（aa）。含有24个c端或n端aa的ena - lp 31aa （dW2Y1）是EBNA2或ena - lp特异性和高效的亲和矩阵。EBNA-LP和EBNA2之间的这些生化相互作用仅在相互作用不稳定时才能在淋巴细胞中协调细胞和病毒基因表达的转录调节；ena - lp c端10aa的缺失稳定了与EBNA2的结合并阻止了共激活。由于EBNA-LPd10主要抑制EBNA-LP与EBNA2的共激活，因此EBNA-LPd10在LCL中的过表达可能有助于评估EBNA-LP共激活在LCL生长或存活中的作用。

项目成果

Shizuko Harada: "Epstein-Barr virus gene expression in latent infection and B-lymphocyte growth transformation."Virus. 52(1). 129-134 (2002)

Shizuko Harada：“Epstein-Barr病毒潜伏感染中的基因表达和B淋巴细胞生长转化。”病毒。

C-W.Peng, Y.Xue, B.Zhao, E.Johanssen, E.Kieff, S.Harada: "Direct interactions between Epstein-Barr virus leader protein (EBNALP) and the EBNA2 acidic domain underlie cooperative transcriptional regulation."Proc.Natl.Acad.Sci.USA. 101(4). 1033-1038 (2004)

C-W.Peng、Y.Xue、B.Zhao、E.Johanssen、E.Kieff、S.Harada：“Epstein-Barr 病毒前导蛋白 (EBNALP) 和 EBNA2 酸性结构域之间的直接相互作用是协同转录调控的基础。”Proc。

T.Nabeshima, A.Mori, T.Kozaki, Y.Iwata, O.Hidoh, S.Harada, S.Kasai, D.W.Severson, Y.Kono, T.Tomita: "An amino acid substitution attributable to insecticide-insensitivity of acetylcholinesterase in Japanese encephalitis vector mosquito, Culex tritaeniorync

T.Nabeshima、A.Mori、T.Kozaki、Y.Iwata、O.Hidoh、S.Harada、S.Kasai、D.W.Severson、Y.Kono、T.Tomita：“由于对杀虫剂不敏感而导致的氨基酸取代

T.Nabeshima, A.Mori, T.Kozaki, Y.Iwata, O.Hidoh, S.Harada, S.Kasai, D.W.Severson, Y.Kono, T.Tomita: "An amino acid substitution attributable to insecticide-insensitivity of acetylcholinesterase in Japanese encephalitis vector mosquito, Culex trilaeniorync

T.Nabeshima、A.Mori、T.Kozaki、Y.Iwata、O.Hidoh、S.Harada、S.Kasai、D.W.Severson、Y.Kono、T.Tomita：“由于对杀虫剂不敏感而导致的氨基酸取代

C-W.Peng, Y.Xue, B.Zhao, E.Johanssen, E.Kieff, S.Harada: "Direct interactions between Epstein-Barr virus leader protein (EBNALP) and the EBNA2 acidic domain underlie cooperative transcriptional regulation."Proc.Natl.Acad.Sci.USA. 101. 1033-1038 (2004)

C-W.Peng、Y.Xue、B.Zhao、E.Johanssen、E.Kieff、S.Harada：“Epstein-Barr 病毒前导蛋白 (EBNALP) 和 EBNA2 酸性结构域之间的直接相互作用是协同转录调控的基础。”Proc。