Gómez-Puertas P, Albo C, Pérez-Pastrana E, Vivo A, Portela A
Centro Nacional de Biología Fundamental, Instituto de Salud Carlos III, Majadahonda 28220, Madrid, Spain.
J Virol. 2000 Dec;74(24):11538-47. doi: 10.1128/jvi.74.24.11538-11547.2000.
To get insights into the role played by each of the influenza A virus polypeptides in morphogenesis and virus particle assembly, the generation of virus-like particles (VLPs) has been examined in COS-1 cell cultures expressing, from recombinant plasmids, different combinations of the viral structural proteins. The presence of VLPs was examined biochemically, following centrifugation of the supernatants collected from transfected cells through sucrose cushions and immunoblotting, and by electron-microscopic analysis. It is demonstrated that the matrix (M1) protein is the only viral component which is essential for VLP formation and that the viral ribonucleoproteins are not required for virus particle formation. It is also shown that the M1 protein, when expressed alone, assembles into virus-like budding particles, which are released in the culture medium, and that the recombinant M1 protein accumulates intracellularly, forming tubular structures. All these results are discussed with regard to the roles played by the virus polypeptides during virus assembly.
为深入了解甲型流感病毒的每种多肽在形态发生和病毒粒子组装中所起的作用,研究人员在COS - 1细胞培养物中检测了病毒样颗粒(VLP)的产生情况。在该细胞培养物中,通过重组质粒表达病毒结构蛋白的不同组合。从转染细胞收集的上清液经蔗糖垫层离心、免疫印迹以及电子显微镜分析后,对VLP的存在情况进行了生化检测。结果表明,基质(M1)蛋白是VLP形成所必需的唯一病毒成分,而病毒核糖核蛋白并非病毒粒子形成所必需。研究还表明,单独表达的M1蛋白可组装成病毒样出芽颗粒,并释放到培养基中,重组M1蛋白在细胞内积累,形成管状结构。针对病毒多肽在病毒组装过程中所起的作用,对所有这些结果进行了讨论。