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FUNCTIONAL ROBES OF HEPATITIS A VIRUS NONSTRUCTURAL PROTEINS IN SIGNAL TRANSDUCTION

甲型肝炎病毒非结构蛋白在信号转导中的功能

基本信息

批准号：
12670458
负责人：
YOKOSUKA Osamu
金额：
$ 1.98万
依托单位：
CHIBA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

项目摘要

Hepatitis A virus (HAV) infection is still a major problem worldwide. The infection does not induce any visible cytopathic effects, and there is no evidence that HAV interferes with the macromolecular synthesis of its host cells. Recently, in the hepatitis B and C virus infection, intracellular signals have been reported to be induced. To Clarify the effects of HAV infection, we examined the influence of 9 FLAG-HAV (VP2, VP3, VP1-2A, 2B, 2C, 3A, 3BC, 3C, 3D) proteins on various intracellular signaling pathways. Viral protein expression vectors were cotransfacted into HeLa cells with the reporter plasmids, controlled by a synthetic promoter that contains direct 4 repeats of the cyclic AMP response element (CRE), 5 repeats of the serum response factor (SRF), 7 repeats of the activator protein 1 (AP 1), 5 repeats of the nuclear factor kB (NF-kB), and 5 repeats of the serum response element (SRE), respectively. Cells were harvested 42 hours after transfaction, and luciferase assays were performed. Assays were conducted at least in triplicate, and a viral protein activation twice greater than that of the control was defined as significant. HAV FLAG-VP3 protein significantly activated the SRE-associated signal in HeLa cells at a value 2.2± 0.3 times higher than control. However, FLAG-VP3 protein did not activate the CRE-(1.1±0.1), SRF-(1.6± 0.2), AP-1-(1.1±0.1), NF-kB-(0.7±0.1) associated pathways. The other HAV proteins showed no signal activation in the CRE-, SRF-, AP-1-, NF-kB-, and SRE-associated pathways. We found that HAV VP3 protein activated the SRE-associated signal, intracellular signaling pathways associated with cell proliferation, differentiation, in HeLa cells.

甲型肝炎病毒（HAV）感染仍然是世界范围内的主要问题。感染不引起任何可见的细胞病变效应，也没有证据表明HAV干扰其宿主细胞的大分子合成。最近，据报道在B和C型肝炎病毒感染中，细胞内信号被诱导。为了阐明HAV感染的影响，我们检测了9种FLAG-HAV（VP 2，VP 3，VP 1 -2A，2B，2C，3A，3BC，3C，3D）蛋白对各种细胞内信号传导途径的影响。将病毒蛋白表达载体与报告质粒共转染到HeLa细胞中，所述报告质粒由合成启动子控制，所述合成启动子含有环AMP反应元件（CRE）的直接4个重复、血清反应因子（SRF）的5个重复、激活蛋白1（AP 1）的7个重复、核因子kB（NF-kB）的5个重复和血清反应元件（SRE）的5个重复，分别在转染后42小时收获细胞，并进行荧光素酶测定。试验至少进行三次，病毒蛋白活化比对照高两倍定义为显著。HAV FLAG-VP 3蛋白显著激活HeLa细胞中的SRE相关信号，其值是对照的2.2± 0.3倍。FLAG-VP 3蛋白对CRE-（1.1±0.1），SRF-（1.6± 0.2），AP-1-（1.1±0.1），NF-kB-（0.7±0.1）通路无激活作用。其他HAV蛋白在CRE-、SRF-、AP-1-、NF-kB-和SRE-相关通路中未显示信号激活。我们发现HAV VP 3蛋白激活了HeLa细胞中与细胞增殖、分化相关的细胞内信号通路SRE相关信号。