Ohgimoto S, Bando H, Kawano M, Okamoto K, Kondo K, Tsurudome M, Nishio M, Ito Y
Department of Microbiology, Mie University School of Medicine, Japan.
Virology. 1990 Jul;177(1):116-23. doi: 10.1016/0042-6822(90)90465-4.
We cloned and sequenced the cDNAs against genomic RNA and mRNA for phosphoprotein (P) of human parainfluenza type 2 virus (PIV-2). cDNA clone from genomic RNA was 1439 nucleotides in length excluding poly(A) and was found to have two small open reading frames encoding proteins of 233 and 249 amino acids. Two different mRNA cDNA clones were obtained; that is, one mRNA contained a smaller reading frame coding 225 amino acids, V protein, and the other mRNA contained a larger reading frame coding 395 amino acids, P protein. Both mRNAs had G cluster in coding frame. The former mRNA contained seven G residues, and two extra G residues were inserted in the latter mRNA. Ten cDNA clones from the genomic RNA were identical and were composed of seven G residues, indicating that genomes analyzed here were a homogeneous population. Therefore, V protein is encoded by faithfully copied mRNA and P protein is translated from mRNA in which two additional G residues are nontemplately inserted immediately after seven genomically encoded G residues. The V and P proteins are amino coterminal proteins and have different C termini. The C terminus of V protein is cysteine-rich and bears some resemblance to metal-binding protein of the zinc finger-type motif. P protein sequence of PIV-2 showed high homologies with SV 5 (40.4%) and mumps virus (35.5%), and a moderate homology with Newcastle disease virus (20.6%). On the other hand, very little homology was found between PIV-2 and other paramyxoviruses including Sendai virus, PIV-3, and measles virus. The cysteine-rich region in V protein was found to be highly conserved in PIV-2, SV 5, and measles virus, suggesting that V protein of paramyxoviruses plays important roles in transcription and/or replication. The predicted cysteine-rich V protein was detected in virus-infected cells using antiserum directed against an oligopeptide specific for the predicted V polypeptide.
我们针对人副流感2型病毒(PIV-2)的磷蛋白(P),克隆了与基因组RNA和mRNA对应的cDNA并进行测序。来自基因组RNA的cDNA克隆长度为1439个核苷酸(不包括聚腺苷酸),发现有两个小开放阅读框,分别编码233和249个氨基酸的蛋白质。获得了两个不同的mRNA cDNA克隆;也就是说,一个mRNA包含一个编码225个氨基酸的较小阅读框,即V蛋白,另一个mRNA包含一个编码395个氨基酸的较大阅读框,即P蛋白。两个mRNA在编码框中都有G簇。前一个mRNA包含七个G残基,后一个mRNA中插入了两个额外的G残基。来自基因组RNA的十个cDNA克隆是相同的,由七个G残基组成,表明这里分析的基因组是一个同质群体。因此,V蛋白由忠实复制的mRNA编码,P蛋白则从mRNA翻译而来,其中在七个基因组编码的G残基之后立即非模板性地插入了两个额外的G残基。V蛋白和P蛋白是氨基共末端蛋白,但C末端不同。V蛋白的C末端富含半胱氨酸,与锌指型基序的金属结合蛋白有一些相似之处。PIV-2的P蛋白序列与SV 5(40.4%)和腮腺炎病毒(35.5%)显示出高度同源性,与新城疫病毒(20.6%)有中等同源性。另一方面,在PIV-2与包括仙台病毒、PIV-3和麻疹病毒在内的其他副粘病毒之间发现的同源性非常低。发现V蛋白中富含半胱氨酸的区域在PIV-2、SV 5和麻疹病毒中高度保守,这表明副粘病毒的V蛋白在转录和/或复制中起重要作用。使用针对预测的V多肽特异性寡肽的抗血清,在病毒感染的细胞中检测到了预测的富含半胱氨酸的V蛋白。