Functions of the mutated NS1 protein of influenza B virus which has a long caroxyl terminal deletion.

具有长羧基末端缺失的乙型流感病毒 NS1 蛋白突变蛋白的功能。

• 批准号: 02670196
• 负责人: TOBITA Kiyotake
• 金额: $ 1.47万
• 依托单位: Jichi Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02670196/
• 关键词: influenza B virus; NS gene; nucleotide sequence; deletion mutation; NS1 protein; cytopathic effect; インフルエンザウイルス; 変異体; RNA合成; CPE

By hetero typic crosses of influenza A and B viruses, we isolated 3 mutants of influenza virus B/Yamagata, which carried deletion mutations in their NS genes, namely, clone 201, AUB2-253 and AWBY-234. Due to a frame-shift downstream, all of them expressed greatly truncated NS1 protein which lacked more than half of the carboxyl terminal portion of the molecule, leaving NS2 intact. To delineate the functions of the NS gene of influenza virus unequivocally, we first prepared a reassortant virus between clone 201 and wild type B/Lee/40, in which only the mutated NS gene of clone 201 was inserted in place of that of B/Lee. As a control, we also prepared a. control reassortant between wild type, Lee/40 and wild type B/Yamagata, in which only the NS gene was substituted with the wild type NS gene of Yamagata. Comparative studies of the two reassortants revealed the following.(1) A large carboxyl terminal deletion of the NSI protein of influenza virus greatly accelerates cell sestruction.(2) Such a deletion mutation does not affect viral protein synthesis.(3) Carboxyl terminal deletion of the NS1 protein alters the mode of VRNA synthesis.The presence of a large amount of subgenomic RNAs within the virion and the occurrence of multiplicity reactivation suggested that a deletion in NS1 protein inhibited efficient elongation of the nascent viral RNA chains.

通过甲型和乙型流感病毒异型杂交，我们分离出3个B/Yamagata流感病毒突变体，其NS基因携带缺失突变，即克隆201、AUB2-253和AWBY-234。由于下游移码，它们都表达了大大截短的 NS1 蛋白，该蛋白缺乏分子一半以上的羧基末端部分，而 NS2 保持完整。为了明确描述流感病毒NS基因的功能，我们首先制备了克隆201和野生型B/Lee/40之间的重配病毒，其中仅插入克隆201的突变NS基因代替B/Lee的NS基因。作为对照，我们还准备了一个。野生型Lee/40和野生型B/Yamagata之间的对照重配，其中仅NS基因被Yamagata野生型NS基因取代。两种重配体的比较研究揭示了以下几点：（1）流感病毒NSI蛋白的羧基末端大缺失大大加速了细胞的构建。（2）这种缺失突变并不影响病毒蛋白的合成。（3）NS1蛋白的羧基末端缺失改变了VRNA的合成方式。大量亚基因组的存在 病毒颗粒内的 RNA 和多重重新激活的发生表明 NS1 蛋白的缺失抑制了新生病毒 RNA 链的有效延长。

项目成果

M. Tashiro, I. James, S. Karri, K. Wahn, K. Tobita, H. -D. Klenk, R. Rott and J. Seto: "Pneumotropic revertants derived from a pantropic mutant, F1-R, of Sendai virus." Virology. 184. 227-234 (1991)

M. Tashiro、I. James、S. Karri、K. Wahn、K. Tobita、H. -D。

M.Tashiro: "Pneumotropic revertants derived from a pantropic mutant,F1-R,of Sendai virus." Virology. 184. 227-234 (1991)

M.Tashiro：“嗜肺回复体源自仙台病毒的泛嗜性突变体 F1-R。”

K.Tobita: "Nucleotide sequence and some biological properties of the NS gene of a newly isolated influenza B virus mutant which has a long carboxyterminal deletion in the NS1 protein." Virology. 174. 314-319 (1990)

K.Tobita：“新分离的乙型流感病毒突变体的 NS 基因的核苷酸序列和一些生物学特性，该突变体在 NS1 蛋白中具有长羧基末端缺失。”

T. Odagiri, K. Tobita and M. Tashiro: "Synthesis of the NS2 nonstructural protein messenger RNA of influenza viruses occurs in the absence of viral protein synthesis." Archives of Virology. 184. 227-234 (1991)

T. Odagiri、K. Tobita 和 M. Tashiro：“流感病毒 NS2 非结构蛋白信使 RNA 的合成是在没有病毒蛋白合成的情况下进行的。”

T.Odagiri: "Synthesis of the NS2 nonstructural protein messenger RNA of influenza viruses occurs in the absence of viral protein synthsis." Archives of Virology. 120. 281-288 (1991)

T.Odagiri：“流感病毒 NS2 非结构蛋白信使 RNA 的合成是在没有病毒蛋白合成的情况下进行的。”