Dewalt P G, Lawson M A, Colonno R J, Semler B L
Department of Microbiology and Molecular Genetics, College of Medicine, University of California, Irvine 92717.
J Virol. 1989 Aug;63(8):3444-52. doi: 10.1128/JVI.63.8.3444-3452.1989.
Cross-species proteolytic processing was demonstrated by the 3C proteinases of human rhinovirus 14 and coxsackievirus B3 on poliovirus-specific polypeptide precursors. Chimeric picornavirus cDNA genomes were constructed in a T7 transcription vector in which the poliovirus 3C coding region was substituted with the corresponding allele from human rhinovirus 14 or coxsackievirus B3. In vitro translation and processing of the polypeptides encoded by the chimeric genomes demonstrated that the proteolytic processing of poliovirus P2 region (nonstructural) proteins could be functionally substituted by the heterologous proteinases. In contrast, the 3C proteinase activities expressed from the chimeric genomes were incapable of recognizing the poliovirus-specific processing sites within the capsid precursor. Since the amino acid sequences flanking and inclusive of the P2 region cleavage sites of the three viruses are not stringently conserved, these results provide evidence for the existence of common conformational determinants necessary for 3C-mediated processing.
人鼻病毒14型和柯萨奇病毒B3型的3C蛋白酶对脊髓灰质炎病毒特异性多肽前体进行了跨物种蛋白水解加工。在T7转录载体中构建了嵌合微小核糖核酸病毒cDNA基因组,其中脊髓灰质炎病毒3C编码区被人鼻病毒14型或柯萨奇病毒B3型的相应等位基因取代。对嵌合基因组编码的多肽进行体外翻译和加工表明,脊髓灰质炎病毒P2区(非结构)蛋白的蛋白水解加工可被异源蛋白酶功能替代。相比之下,嵌合基因组表达的3C蛋白酶活性无法识别衣壳前体内脊髓灰质炎病毒特异性加工位点。由于这三种病毒P2区切割位点两侧及包含该位点的氨基酸序列并非严格保守,这些结果为3C介导的加工所需的共同构象决定簇的存在提供了证据。