Singh M K, Fuller-Pace F V, Buchmeier M J, Southern P J
Department of Immunology, Scripps Clinic and Research Foundation, La Jolla, California 92037.
Virology. 1987 Dec;161(2):448-56. doi: 10.1016/0042-6822(87)90138-3.
The arenavirus genomic L RNA segment represents approximately 70% of the viral genetic material but current understanding of the organization, regulation, and functioning of the viral L products remains limited. Biological studies with reassortant viruses have implicated the L RNA segment in the lethal infection of adult guinea pigs produced by LCMV-WE but no further explanation of the pathogenic process is presently available. We have initiated a detailed molecular analysis of LCMV L products based on construction and characterization of L-specific cDNA clones and synthesis of L-specific hybridization probes. Nucleotide sequencing studies have allowed the derivation of a partial amino acid sequence for a predicted L protein and antisera raised against synthetic peptides have demonstrated an L protein in Western blotting experiments. Using this approach we have identified a single high molecular weight protein (approximately 200,000 Da) in purified virions and in viral ribonucleoprotein complexes extracted from acutely infected tissue culture cells. This L protein is translated from a nonpolyadenylated, genomic complementary L mRNA and potentially represents part or all of the viral RNA-dependent RNA polymerase.
沙粒病毒基因组L RNA片段约占病毒遗传物质的70%,但目前对病毒L产物的组织、调控和功能的了解仍然有限。对重组病毒的生物学研究表明,L RNA片段与LCMV-WE引起的成年豚鼠致死性感染有关,但目前尚无对致病过程的进一步解释。我们基于L特异性cDNA克隆的构建和表征以及L特异性杂交探针的合成,启动了对LCMV L产物的详细分子分析。核苷酸测序研究已推导预测的L蛋白的部分氨基酸序列,针对合成肽产生的抗血清在蛋白质印迹实验中证实了L蛋白的存在。利用这种方法,我们在纯化的病毒粒子以及从急性感染的组织培养细胞中提取的病毒核糖核蛋白复合物中鉴定出一种单一的高分子量蛋白(约200,000 Da)。这种L蛋白由非聚腺苷酸化的基因组互补L mRNA翻译而来,可能代表病毒RNA依赖性RNA聚合酶的部分或全部。