An African swine fever virus ERV1-ALR homologue, 9GL, affects virion maturation and viral growth in macrophages and viral virulence in swine
An African swine fever virus ERV1-ALR homologue, 9GL, affects virion maturation and viral growth in macrophages and viral virulence in swine
复制标题
DOI：
10.1128/jvi.74.3.1275-1285.2000
复制
发表时间：
2000-02-01
影响因子：
5.4
通讯作者：
Rock, DL
中科院分区：
文献类型：
--
作者：
Lewis, T;Zsak, L;Rock, DL
非洲猪瘟病毒(ASFV)基因组包含一个基因9GL，与酵母ERV1和ALR基因相似。ERV1被证明在氧化磷酸化和细胞生长中发挥作用，而ALR具有肝营养活性。9GL编码119个氨基酸的蛋白质，在核苷酸和氨基酸水平上都高度保守。针对谷胱甘肽S-转移酶-9GL融合蛋白制备的兔多克隆抗体，可从感染ASFV马拉维株LIL-20/1(MAL)的巨噬细胞培养物中免疫沉淀出一条14 kDa的蛋白。时程分析和病毒DNA合成抑制实验表明，p14是一种晚期病毒蛋白。MAL的9GL基因缺失突变体(Delta 9GL)，在巨噬细胞中表现出约2log(10)单位的生长缺陷，与回复突变体(9GL-R)或亲本病毒相比，具有小的空斑表型。9GL影响正常的病毒粒子成熟；在Delta 9GL感染的巨噬细胞中，含有无着丝粒类核结构的病毒粒子占所有病毒粒子的90%到99%。Delta 9GL病毒在猪体内的感染明显减弱。与死亡率为100%的9GL-R感染相反，感染Delta 9GL的动物全部存活。除了一些动物的短暂性发热反应外，感染Delta 9GL的动物在临床上保持正常，病毒血症滴度显著降低100至10,000倍。所有先前感染Delta 9GL的猪在随后被致死剂量的亲本MAL攻击时，都存活了下来。因此，ASFV 9GL基因缺失突变体可能被证明是有用的ASF减毒活疫苗。
The African swine fever virus (ASFV) genome contains a gene, 9GL, with similarity to yeast ERV1 and ALR genes. ERV1 has been shown to function in oxidative phosphorylation and in cell growth, while ALR has hepatotrophic activity. 9GL encodes a protein of 119 amino acids and was highly conserved at both nucleotide and amino acid levels among all ASFV field isolates examined. Monospecific rabbit polyclonal antibody produced to a glutathione S-transferase-9GL fusion protein specifically immunoprecipitated a 14-kDa protein from macrophage cell cultures infected with the ASFV isolate Malawi Lil-20/1 (MAL). Time course analysis and viral DNA synthesis inhibitor experiments indicated that p14 was a late viral protein. A 9GL gene deletion mutant of MAL (Delta 9GL), exhibited a growth defect in macrophages of approximately 2 log(10) units and had a small-plaque phenotype compared to either a revertant (9GL-R) or the parental virus. 9GL affected normal virion maturation; virions containing acentric nucleoid structures comprised 90 to 99% of all virions observed in Delta 9GL-infected macrophages. The Delta 9GL virus was markedly attenuated in swine. In contrast to 9GL-R infection, where mortality was 100%, all Delta 9GL-infected animals survived infection. With the exception of a transient fever response in some animals, Delta 9GL-infected animals remained clinically normal and exhibited significant 100- to 10,000-fold reductions in viremia titers. All pigs previously infected with Delta 9GL survived infection when subsequently challenged with a lethal dose of virulent parental MAL. Thus, ASFV 9GL gene deletion mutants may prove useful as live-attenuated ASF vaccines.