Fernholz D, Galle P R, Stemler M, Brunetto M, Bonino F, Will H
Max-Planck-Institut für Biochemie, Martinsried/München, Germany.
Virology. 1993 May;194(1):137-48. doi: 10.1006/viro.1993.1243.
All human hepatitis B viruses characterized so far express three envelope proteins, pre-S1, pre-S2, and HBs, which are believed to function as binding proteins for the cellular receptor, as targets for immune-mediated virus elimination, and in virion morphogenesis and secretion. Here we report the characterization of infectious HBV variant genomes that are unable to express a pre-S2 protein and which were derived from serum of a highly viremic chronic carrier. Direct sequencing of the amplified pre-S region and sequencing of 50 cloned amplified pre-S DNA fragments revealed that in all molecules, in addition to numerous nucleotide changes, there were deletions of the pre-S2 translation initiation codon and three codons 54 nucleotides downstream thereof. No pre-S2 protein and altered pre-S1 proteins were found in the serum of the patient. Cloned infectious HBV DNA genomes having the pre-S region substituted by the variant pre-S region were replication competent in cultured hepatoblastoma cells. Morphologically normal virions were efficiently secreted and were infectious for primary human hepatocyte cultures. These data demonstrate that HBV devoid of pre-S2 protein can occur in vivo as a dominant or exclusive virus population and that expression of the pre-S2 protein is not essential for HBV replication, virion morphogenesis, secretion, or in vitro infectivity.
迄今为止所鉴定的所有人类乙型肝炎病毒均表达三种包膜蛋白,即前S1、前S2和乙肝表面抗原(HBs),据信它们作为细胞受体的结合蛋白、免疫介导的病毒清除靶点以及在病毒体形态发生和分泌过程中发挥作用。在此,我们报告了感染性乙肝病毒(HBV)变异基因组的特征,这些基因组无法表达前S2蛋白,且来源于一名高病毒血症慢性携带者的血清。对扩增的前S区域进行直接测序以及对50个克隆的扩增前S DNA片段进行测序显示,在所有分子中,除了众多核苷酸变化外,前S2翻译起始密码子及其下游54个核苷酸处的三个密码子均有缺失。在该患者的血清中未发现前S2蛋白和改变的前S1蛋白。用变异前S区域替换前S区域的克隆感染性HBV DNA基因组在培养的肝癌细胞中具有复制能力。形态正常的病毒体被有效分泌,并且对原代人肝细胞培养物具有感染性。这些数据表明,缺乏前S2蛋白的HBV可以在体内作为主要或唯一的病毒群体出现,并且前S2蛋白的表达对于HBV复制、病毒体形态发生、分泌或体外感染性并非必需。