Complementation of P37 (F13L gene) knock-out in vaccinia virus by a cell line expressing the gene constitutively
Complementation of P37 (F13L gene) knock-out in vaccinia virus by a cell line expressing the gene constitutively
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DOI：
10.1099/0022-1317-80-2-425
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发表时间：
1999-02-01
影响因子：
3.8
通讯作者：
Blasco, R
中科院分区：
文献类型：
--
作者：
Borrego, B;Lorenzo, MM;Blasco, R
痘苗病毒产生两种不同的感染形式，细胞内成熟病毒 (IMV) 和细胞外包膜病毒 (EEV)，EEV 包膜的获得是通过将 IMV 与跨高尔基体网络 (TGN) 的囊泡包裹而实现的。EEV 包膜中最丰富的蛋白质 P37 是由 F13L 基因编码的 37 kDa 棕榈酰化蛋白质。 P37 位于 EEV 包膜的内侧，并在感染期间积聚在 TGN 中。尽管产生了正常水平的 IMV，但 F13L 基因的缺失会导致包裹过程中的严重缺陷。获得了源自 RK-13 细胞的稳定表达 P37 (RKP37) 的细胞系，并在没有其他病毒多肽的情况下研究了该蛋白质的特性。 RKP37 细胞中产生的 P37 与牛痘感染细胞中产生的 P37 在疏水性和细胞内分布方面有所不同。尽管存在这些差异，RKP37 细胞部分补充了痘苗病毒 P37(-) 突变体的表型缺陷。与正常 RK-13 细胞培养物相比，RKP37 细胞中突变病毒的 EEV 产生和细胞间病毒传播显着增加。用P37(-)病毒感染RKP37细胞显着改变了这些细胞中P37的疏水性和细胞内分布。这些结果表明，确定 P37 的正常棕榈酰化和细胞内定位需要感染环境。
Vaccinia virus produces two different infectious forms, intracellular mature virus (IMV) and extracellular enveloped virus (EEV), Acquisition of the EEV envelope occurs by wrapping of IMV with vesicles of the trans-Golgi network (TGN), The most abundant protein in the envelope of EEV, P37, is a 37 kDa palmitylated protein encoded by the F13L gene. P37 is located in the inner side of the EEV envelope and accumulates in the TGN during infection. Deletion of gene F13L results in a severe defect in the wrapping process, although normal levels of IMV are produced. A cell line, derived from RK-13 cells, was obtained that stably expressed P37 (RKP37), and the properties of the protein were studied in the absence of other viral polypeptides. P37 produced in RKP37 cells differed from P37 produced in vaccinia-infected cells in terms of hydrophobicity and intracellular distribution. Despite these differences, RKP37 cells partially complemented the phenotypic defect of vaccinia virus P37(-) mutants. EEV production and cell-to-cell virus spread by mutant viruses were increased significantly in RKP37 cells when compared to normal RK-13 cell cultures. Infection of RKP37 cells with P37(-) virus substantially altered the hydrophobicity and the intracellular distribution of P37 in those cells. These results indicate the requirement of the infection context for determination of the normal palmitylation and intracellular localization of P37.