Urban S, Breiner K M, Fehler F, Klingmüller U, Schaller H
Zentrum für Molekulare Biologie, Universität Heidelberg, 69120 Heidelberg, Germany.
J Virol. 1998 Oct;72(10):8089-97. doi: 10.1128/JVI.72.10.8089-8097.1998.
Functionally relevant hepadnavirus-cell surface interactions were investigated with the duck hepatitis B virus (DHBV) animal model by using an in vitro infection competition assay. Recombinant DHBV pre-S polypeptides, produced in Escherichia coli, were shown to inhibit DHBV infection in a dose-dependent manner, indicating that monomeric pre-S chains were capable of interfering with virus-receptor interaction. Particle-associated pre-S was, however, 30-fold more active, suggesting that cooperative interactions enhance particle binding. An 85-amino-acid pre-S sequence, spanning about half of the DHBV pre-S chain, was characterized by deletion analysis as essential for maximal inhibition. Pre-S polypeptides from heron hepatitis B virus (HHBV) competed DHBV infection equally well despite a 50% difference in amino acid sequence and a much-reduced infectivity of HHBV for duck hepatocytes. These observations are taken to indicate (i) that the functionality of the DHBV pre-S subdomain, which interacts with the cellular receptor, is determined predominantly by a defined three-dimensional structure rather than by primary sequence elements; (ii) that cellular uptake of hepadnaviruses is a multistep process involving more than a single cellular receptor component; and (iii) that gp180, a cellular receptor candidate unable to discriminate between DHBV and HHBV, is a common component of the cellular receptor complex for avian hepadnaviruses.
利用体外感染竞争试验,通过鸭乙型肝炎病毒(DHBV)动物模型研究了功能相关的嗜肝DNA病毒-细胞表面相互作用。在大肠杆菌中产生的重组DHBV前S多肽以剂量依赖方式抑制DHBV感染,这表明单体前S链能够干扰病毒-受体相互作用。然而,与病毒颗粒相关的前S活性高30倍,这表明协同相互作用增强了病毒颗粒的结合。通过缺失分析确定,跨越DHBV前S链约一半的85个氨基酸的前S序列对于最大抑制至关重要。尽管鹭乙型肝炎病毒(HHBV)的氨基酸序列有50%的差异,且其对鸭肝细胞的感染性大大降低,但其前S多肽对DHBV感染的竞争效果同样良好。这些观察结果表明:(i)与细胞受体相互作用的DHBV前S亚结构域的功能主要由确定的三维结构而非一级序列元件决定;(ii)嗜肝DNA病毒的细胞摄取是一个多步骤过程,涉及多个细胞受体成分;(iii)gp180作为一种无法区分DHBV和HHBV的细胞受体候选物,是禽类嗜肝DNA病毒细胞受体复合物的共同成分。