A 14K ENVELOPE PROTEIN OF VACCINIA VIRUS WITH AN IMPORTANT ROLE IN VIRUS HOST-CELL INTERACTIONS IS ALTERED DURING VIRUS PERSISTENCE AND DETERMINES THE PLAQUE SIZE PHENOTYPE OF THE VIRUS
A 14K ENVELOPE PROTEIN OF VACCINIA VIRUS WITH AN IMPORTANT ROLE IN VIRUS HOST-CELL INTERACTIONS IS ALTERED DURING VIRUS PERSISTENCE AND DETERMINES THE PLAQUE SIZE PHENOTYPE OF THE VIRUS
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DOI：
10.1016/0042-6822(87)90481-8
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发表时间：
1987-08-01
期刊：
影响因子：
3.7
通讯作者：
ESTEBAN, M
中科院分区：
文献类型：
--
作者：
DALLO, S;RODRIGUEZ, JF;ESTEBAN, M
人们对痘病毒的遗传变异和减毒或毒力现象知之甚少。我们从未经处理和干扰素 (IFN) 处理的持续感染的 Friend 红白血病 (FEL) 细胞中鉴定出痘苗病毒突变体，其病毒结构蛋白的大小发生了重大变化。该蛋白是用特异性单克隆抗体 (mAbC3) 包裹的病毒的一部分。在还原条件下，该蛋白在野生型病毒中的分子量约为 14,000 (14K) Da，但在未经处理的持续感染细胞的突变体中分子量增加了 1.5 K，在 IFN 处理的持续感染细胞的突变体中分子量增加了约 0.5 K。在非还原条件下，该蛋白质形成共价连接的寡聚体，其大小在野生型和突变病毒之间也不同。使用兔抗痘苗血清通过二维 SDS-PAGE 分析的免疫印迹评估 14K 蛋白引发体液免疫反应。仅在未经处理的持续感染细胞的突变体中发现了两种具有不同等电点的 14K 蛋白分子形式。蛋白质修饰是病毒群体中 DNA 序列改变的结果，因为在用克隆的 14K 编码基因进行标记拯救后，15.5K 蛋白质可以恢复为 14K。我们提供的直接证据表明，14K 包膜蛋白大小的变化是造成这些变体的小噬斑大小表型的原因。根据我们之前的研究（J.F. Rodriquez, R. Janeczko, and M. Esteban, 1985, J. Virol. 56, 352-356；J. F. Rodriquez, E. Paez, and M. Esteban, 1987, J. Virol. 61,395-404）以及此处描述的研究，我们确定 14K 蛋白在病毒与宿主细胞的相互作用。这是因为该蛋白是病毒包膜的一部分，具有高抗原性，引发中和抗体，在细胞融合中的病毒渗透中发挥作用，参与病毒噬斑大小表型，并且在正痘病毒中高度保守。此外，由于 14K 蛋白改变且大小增加了 1.5K 的突变体被减毒（S. Dallo 和 M. Esteban，1987，1987，Virology 159-422），这表明该蛋白的修饰形式有助于降低痘苗病毒的毒力。
The phenomenon of genetic variability and attenuation or virulence of poxviruses is poorly understood. We have identified mutants of vaccinia virus from untreated and interferon (IFN)-treated persistently infected Friend erythroleukemia (FEL) cells that have major alterations in the size of a virus structural protein. This protein is part of the virus enveloped as documented with specific monoclonal antibody (mAbC3). This protein, under reducing conditions, has a molecular weight of about 14,000 (14K) Da in wild-type virus but gained 1.5 K in mutants from untreated, persistently infected cells and about 0.5K in mutants from IFN-treated persistently infected cells. Under nonreducing conditions, this protein forms covalently linked oligomers which also differ in size between wild-type and mutant viruses. The 14K protein elicits humoral immune response as assessed by immunoblots of two-dimensional SDS-PAGE analysis using rabbit anti-vaccinia serum. Two molecular forms of the 14K protein with different isoelectric points were found only in mutants from untreated persistently infected cells. Protein modifications were the result of DNA sequence alterations in the virus population since the 15.5K protein could be reverted to 14K after marker-rescue with the cloned 14K encoding gene. We provide direct evidence that changes in size of the 14K envelope protein are responsible for the small plaque size phenotype of these variants. From our previous studies (J.F. Rodriquez, R. Janeczko, and M. Esteban, 1985, J. Virol. 56, 352-356; J. F. Rodriquez, E. Paez, and M. Esteban, 1987, J. Virol. 61,395-404) and those described here, we established that the 14K protein plays an important role in virus-host cell interactions. This is because this protein is part of the virus envelope, is higly antigenic, elicits neutralizing antibodies, has a role in virus penetration acting in cell fusion, is involved in the virus plaque size phenotype and is highly conserved among Orthopoxviruses. In addition, because mutants with altered 14K protein that have gained 1.5K in size are attenuated (S. Dallo and M. Esteban, 1987, 1987, Virology 159-422), it suggests that a modified form of this protein contributes to the decrease of the virulence of vaccinia virus.