Onodera S, Cardamone J J, Phillips B A
J Virol. 1986 May;58(2):610-8. doi: 10.1128/JVI.58.2.610-618.1986.
Highly purified 14S subunit particles were obtained from alkali-dissociated poliovirus type 1 procapsids (naturally occurring empty capsids in poliovirus-infected cells) to compare their morphological and biophysical properties with those of naturally occurring 14S particles. Procapsid-derived 14S particles (PC-14S), like naturally occurring 14S particles, were capable of self-assembly into an empty shell in buffer or extracts from uninfected cells. These empty capsids always exhibited pIs more acidic than those of procapsids but were themselves distinguishable by their respective pIs. Nevertheless, if PC-14S or naturally occurring 14S particles were incubated with extracts made from poliovirus-infected cells, procapsidlike empty shells were formed. This clearly showed that the 14S particle, however obtained, possesses the information to form an empty shell of correct dimensions but of improper conformation, unless a factor present in poliovirus-infected cells is present. With the electron microscope, the PC-14S subunit frequently was seen as a pentagonal structure with a diameter of 20.4 +/- 1.4 nm, a size somewhat larger than expected for a subunit composing 1/12th of the poliovirus surface. Upon self-assembly in vitro, the empty shell formed exhibited a diameter of 29 +/- 1 nm and a wall thickness of ca. 6 to 7 nm. It was necessary to avoid CsCl banding of procapsids in their preparation as this treatment altered both their pI and their sensitivity to alkali dissociation into 14S subunits. The relevance of these findings to the nature and role of procapsids and the requirement for a morphopoietic factor in poliovirus morphogenesis is discussed.
从1型脊髓灰质炎病毒前衣壳(脊髓灰质炎病毒感染细胞中天然存在的空衣壳)经碱解离获得了高度纯化的14S亚基颗粒,以将其形态学和生物物理特性与天然存在的14S颗粒的特性进行比较。源自前衣壳的14S颗粒(PC - 14S)与天然存在的14S颗粒一样,能够在缓冲液或未感染细胞的提取物中自组装成空壳。这些空衣壳的等电点总是比前衣壳的更酸性,但它们自身可通过各自的等电点区分开来。然而,如果将PC - 14S或天然存在的14S颗粒与脊髓灰质炎病毒感染细胞制备的提取物一起孵育,就会形成前衣壳样的空壳。这清楚地表明,无论如何获得的14S颗粒都拥有形成尺寸正确但构象不正确的空壳的信息,除非存在脊髓灰质炎病毒感染细胞中存在 的一种因子。用电子显微镜观察,PC - 14S亚基经常被视为直径为20.4±1.4 nm的五边形结构,该尺寸略大于构成脊髓灰质炎病毒表面1/12的亚基预期尺寸。在体外自组装时,形成的空壳直径为29±l nm,壁厚约为6至7 nm。在前衣壳的制备过程中必须避免用CsCl进行区带离心,因为这种处理会改变它们的等电点以及它们对碱解离成14S亚基的敏感性。讨论了这些发现与前衣壳的性质和作用以及脊髓灰质炎病毒形态发生中形态发生因子的需求的相关性。