Ludert J E, Feng N, Yu J H, Broome R L, Hoshino Y, Greenberg H B
Department of Medicine and Microbiology and Immunology, Stanford University School of Medicine, California 94305, USA.
J Virol. 1996 Jan;70(1):487-93. doi: 10.1128/JVI.70.1.487-493.1996.
To identify the rotavirus protein which mediates attachment to cells in culture, viral reassortants between the simian rotavirus strain RRV and the murine strains EHP and EW or between the simian strain SA-11 and the human strain DS-1 were isolated. These parental strains differ in the requirement for sialic acid to bind and infect cells in culture. Infectivity and binding assays with the parental and reassortant rotaviruses indicate that gene 4 encodes the rotavirus protein which mediates attachment to cells in culture for both sialic acid-dependent and -independent strains. Using ligated intestinal segments of newborn mice and reassortants obtained between the murine strain EW and RRV, we developed an in vivo infectivity assay. In this system, the infectivity of EW was not affected by prior treatment of the enterocytes with neuraminidase, while neuraminidase treatment reduced the infectivity of a reassortant carrying gene 4 from RRV on an EW background more than 80% relative to the controls. Thus, VP4 appears to function as the cell attachment protein in vivo as well as in vitro.
为了鉴定介导轮状病毒在细胞培养中附着的蛋白质,我们分离了猿猴轮状病毒RRV株与鼠类EHP和EW株之间,或猿猴SA - 11株与人类DS - 1株之间的病毒重配株。这些亲代毒株在结合和感染细胞培养物时对唾液酸的需求方面存在差异。对亲代和重配轮状病毒进行的感染性和结合试验表明,基因4编码的轮状病毒蛋白介导了唾液酸依赖性和非依赖性毒株在细胞培养中的附着。利用新生小鼠的结扎肠段以及鼠类EW株和RRV株之间获得的重配株,我们开发了一种体内感染性试验。在这个系统中,EW的感染性不受神经氨酸酶预先处理肠上皮细胞的影响,而神经氨酸酶处理使携带RRV基因4的重配株在EW背景下的感染性相对于对照降低了80%以上。因此,VP4似乎在体内和体外均作为细胞附着蛋白发挥作用。