SEQUENCE-ANALYSIS OF THE VIRAL CORE PROTEIN AND THE MEMBRANE-ASSOCIATED PROTEIN-V1 AND PROTEIN-NV2 OF THE FLAVIVIRUS WEST NILE VIRUS AND OF THE GENOME SEQUENCE FOR THESE PROTEINS
SEQUENCE-ANALYSIS OF THE VIRAL CORE PROTEIN AND THE MEMBRANE-ASSOCIATED PROTEIN-V1 AND PROTEIN-NV2 OF THE FLAVIVIRUS WEST NILE VIRUS AND OF THE GENOME SEQUENCE FOR THESE PROTEINS
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DOI：
10.1016/0042-6822(85)90156-4
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发表时间：
1985-01-01
期刊：
影响因子：
3.7
通讯作者：
WENGLER, G
中科院分区：
文献类型：
--
作者：
CASTLE, E;NOWAK, T;WENGLER, G
除了病毒核心蛋白V2之外，细胞相关黄病毒还含有2种膜蛋白V3和NV2，而细胞外病毒确实含有V2蛋白和2种膜蛋白V3和V1。由于在感染的细胞中不能检测到V1蛋白，因此表明V1是在病毒从细胞释放期间通过蛋白水解裂解由NV2产生的。从黄病毒西尼罗病毒中分离病毒结构蛋白V1、V2和NV2，并测定它们的氨基末端氨基酸序列和衍生自这些蛋白的肽的氨基酸序列。将病毒基因组的部分转录成互补DNA，并克隆互补DNA并测序。V1、V2和NV2的蛋白质结构分析以及V3氨基末端序列的测定（数据未显示）允许鉴定编码结构蛋白V2、NV2和V1的核苷酸区域。该核苷酸序列的一级结构见本报告。数据显示，病毒核心蛋白V2的氨基末端之后分别是蛋白NV2、V1和V3的氨基末端。这些数据首次确定了迄今为止鉴定的黄病毒所有结构蛋白的确切顺序。这些数据有力地支持了上述假设，即V1是通过蛋白水解裂解从NV2衍生的，并且进一步表明V1代表NV2的非糖基化羧基末端部分，其含有将NV2锚在病毒膜中的那些序列。提出了一个工作假设，其中2个物种。在细胞酶中，去除信号序列的信号酶和参与切割一对碱性氨基酸后的多聚蛋白的酶确实从42 S基因组RNA翻译期间合成的生长肽链产生蛋白质V2、NV 2和V1，所述42 S基因组RNA作为这些蛋白质的mRNA发挥作用。
Cell-associated flaviviruses contain the 2 membrane proteins V3 and NV2 besides the viral core protein V2 whereas extracellular viruses do contain V2 protein and the 2 membrane proteins V3 and V1. Since the V1 protein could not be detected in infected cells it was suggested that V1 is generated from NV2 by proteolytic cleavage during the release of virus from cells. The viral structural proteins V1, V2, and NV2 were isolated from the flavivirus West Nile virus and their amino-terminal amino acid sequences and amino acid sequences of peptides derived from these proteins were determined. Parts of the viral genome were transcribed into complementary DNA and complementary DNA was cloned and sequenced. The analyses of the protein structure of V1, V2, and NV2 together with the determination of the amino-terminal sequence of V3 (data not shown) have allowed for the identification of the nucleotide region coding for the structural proteins V2, NV2, and V1. The primary structure of this nucleotide sequence is presented in this report. The data show that the amino terminus of the viral core protein V2 is followed by the amino termini of the proteins NV2, V1 and V3, respectively. These data for the 1st time identify the exact order of all structural proteins of a flavivirus identified so far. These data strongly support the above-mentioned hypothesis that V1 is derived from NV2 by proteolytic cleavage and furthermore indicate that V1 represents the nonglycosylated carboxy-terminal part of NV2 which contains those sequences which anchor NV2 in the viral membrane. A working hypothesis is presented in which 2 spp. of cellular enzymes, signalase(s) removing signal sequences and enzymes involved in cleaving polyproteins after a pair of basic amino acids, do generate the proteins V2, NV2 and V1 from the growing peptide chain synthesized during translation of the 42 S genome RNA which functions as mRNA for these proteins.