Seifer M, Standring D N
Hormone Research Institute, University of California, San Francisco 94143-0534.
Virology. 1993 Sep;196(1):70-8. doi: 10.1006/viro.1993.1455.
The p21.5 capsid or core protein of hepatitis B virus carries two distinct classes of epitopes. Core (HBc) epitopes are found exclusively on the surface of the 28-nm viral icosahedral capsids or core particles, while HBe epitopes are normally expressed only by subparticulate forms of the core protein. Recent studies have suggested that a "particulate" form of HBe is expressed on the surface of capsid particles assembled from p17, a truncated core protein that lacks the carboxy-terminal protamine-like region of p21.5 and hence the ability to bind and encapsidate RNA. In this report we have used epitope-specific ELISAs in conjunction with capsids assembled from a series of carboxy-terminally truncated core proteins to address the mechanistic basis for particulate HBe. Specifically, we sought to test the idea that particulate HBe expression might be linked to the loss of RNA binding. However, our results strongly suggest that expression of HBe by mutant core particles is a result of their intrinsic instability which increases sharply when RNA binding is lost. We show that core particles assembled from mutant core proteins lacking Cys residues also express HBe, again because of capsid instability. We report mild conditions that can induce the dissociation of the mutant capsids and discuss our findings in terms of the factors that control capsid stability.
乙肝病毒的p21.5衣壳或核心蛋白带有两类不同的表位。核心(HBc)表位仅存在于28纳米病毒二十面体衣壳或核心颗粒的表面,而HBe表位通常仅由核心蛋白的亚颗粒形式表达。最近的研究表明,一种“HBe颗粒”形式在由p17组装的衣壳颗粒表面表达,p17是一种截短的核心蛋白,缺少p21.5的羧基末端鱼精蛋白样区域,因此缺乏结合和包裹RNA的能力。在本报告中,我们使用表位特异性酶联免疫吸附测定(ELISA),结合由一系列羧基末端截短的核心蛋白组装的衣壳,来探讨颗粒状HBe的机制基础。具体而言,我们试图验证颗粒状HBe表达可能与RNA结合丧失有关这一观点。然而,我们的结果强烈表明,突变核心颗粒表达HBe是其内在不稳定性的结果,当RNA结合丧失时,这种不稳定性会急剧增加。我们表明,由缺乏半胱氨酸残基的突变核心蛋白组装的核心颗粒也表达HBe,同样是由于衣壳不稳定性。我们报告了能够诱导突变衣壳解离的温和条件,并根据控制衣壳稳定性的因素讨论了我们的发现。