Myristylation of the large surface protein is required for hepatitis B virus in vitro infectivity
Myristylation of the large surface protein is required for hepatitis B virus in vitro infectivity
复制标题
DOI：
10.1006/viro.1996.0209
复制
发表时间：
1996-04-15
期刊：
影响因子：
3.7
通讯作者：
Galle, PR
中科院分区：
文献类型：
--
作者：
Bruss, V;Hagelsten, J;Galle, PR
有包膜的B型肝炎病毒（HBV）的大表面蛋白（L）在甘氨酸2处被肉豆蔻酰化。为了研究这种脂肪酸部分是否是HBV感染性所必需的，我们使用了一个点突变体，其中肉豆蔻基受体突变为无功能的丙氨酸。通过基因组DNA构建体的转染，在人肝癌细胞系中表达突变病毒和野生型对照。通过免疫沉淀病毒粒子的内源性聚合酶活性测定，两种菌株分泌的病毒粒子数量相当。L的N-末端表位在病毒体表面的介绍是不受突变的影响。为了测试该突变病毒的感染性，将原代人肝细胞与转染细胞的培养基一起孵育。通过敏感的基于PCR的技术，将感染后产生的共价环状封闭的HBV DNA分子与接种病毒粒子的开放环状DNA基因组区分开来。实验证明野生型是感染性的，但肉豆蔻酸阴性突变体不是。这反映了同源鸭肝炎B病毒突变体的表型，尽管该病毒和HBV的N-末端L蛋白结构域没有显示出一级序列同源性。(C)出版社：Academic Press，Inc.
The large surface protein (L) of the enveloped hepatitis B virus (HBV) is myristylated at glycine 2. To investigate whether this fatty acid moiety is required for HBV infectivity we made use of a point mutant in which the myristyl acceptor was mutated to a nonfunctional alanine. The mutant virus and a wild-type control were expressed in a human hepatoma cell line by transfection of genomic DNA constructs. Comparable amounts of virions were secreted by both strains as measured by the endogenous polymerase activity of immunoprecipitated virions. The presentation of an N-terminal epitope of L on the virion surface was not influenced by the mutation. To test the infectivity of this mutant virus primary human hepatocytes were incubated with the media of transfected cells. The covalently circular closed HBV DNA molecules generated after infection were discriminated from the open circular DNA genomes of inoculated virions by a sensitive PCR-based technique. The experiments demonstrated that the wild type was infectious but not the myristate negative mutant. This reflects the phenotype of an homologous duck hepatitis B virus mutant although the N-terminal L protein domains of this virus and of HBV show no primary sequence homology. (C) 1996 Academic Press, Inc.