Spriggs M K, Collins P L
J Virol. 1986 Sep;59(3):646-54. doi: 10.1128/JVI.59.3.646-654.1986.
cDNA clones of mRNAs for the major nucleocapsid protein (NP), the nucleocapsid P protein plus the nonstructural C protein (P+C), and the matrix protein (M) of human parainfluenza virus type 3 (PF3) were identified by hybrid arrest and hybrid selection of in vitro translation. Previously, cDNA clones were identified and sequenced for the hemagglutinin-neuraminidase glycoprotein (HN) and the fusion glycoprotein (F) mRNAs (N. Elango, J. E. Coligan, R. C. Jambou, and S. Venkatesan, J. Virol. 57:481-489, 1986; M. K. Spriggs, R. A. Olmsted, S. Venkatesan, J. E. Coligan, and P. L. Collins, Virology 152:241-251, 1986). Synthetic oligonucleotides, designed from nucleotide sequences of the cDNAs, were used to direct dideoxynucleotide sequencing of gene junctions in PF3 genomic RNA (vRNA). From sequencing of vRNA, a sixth viral gene was detected and identified as the large nucleocapsid protein (L) gene by hybridization of a synthetic oligonucleotide to intracellular PF3 mRNAs separated by gel electrophoresis. The order of the six PF3 genes on vRNA was 3'-NP-P+C-M-F-HN-L-5'. The five intergenic regions consisted of the trinucleotide 3'-GAA. The PF3 genes initiated with semiconserved 10-nucleotide gene-start sequences and terminated with semiconserved 12-nucleotide gene-end sequences. The M gene terminated with an aberrant gene-end sequence; analysis of intracellular mRNA showed that this aberrant sequence correlated with a disproportionately high accumulation of readthrough mRNA. These studies showed that PF3 encodes six unique mRNAs (NP, P+C, M, F, HN, and L) that encode seven proteins (NP, P, C, M, F, HN, and L) and provided evidence of a close relationship between PF3 and Sendai (murine parainfluenza type 1) viruses.
通过体外翻译的杂交捕获和杂交筛选，鉴定出了人副流感病毒3型（PF3）主要核衣壳蛋白（NP）、核衣壳P蛋白加非结构C蛋白（P+C）以及基质蛋白（M）的mRNA的cDNA克隆。此前，已鉴定出血凝素-神经氨酸酶糖蛋白（HN）和融合糖蛋白（F）mRNA的cDNA克隆并进行了测序（N. 埃兰戈、J. E. 科利根、R. C. 詹布和S. 文卡特桑，《病毒学杂志》57:481 - 489, 1986；M. K. 斯普里格斯、R. A. 奥尔姆斯特德、S. 文卡特桑、J. E. 科利根和P. L. 柯林斯，《病毒学》152:241 - 251, 1986）。根据cDNA的核苷酸序列设计的合成寡核苷酸，用于指导PF3基因组RNA（vRNA）中基因接头的双脱氧核苷酸测序。通过对vRNA的测序，检测到了第六个病毒基因，并通过合成寡核苷酸与经凝胶电泳分离的细胞内PF3 mRNA杂交，将其鉴定为大核衣壳蛋白（L）基因。vRNA上六个PF3基因的顺序为3'-NP-P+C-M-F-HN-L-5'。五个基因间隔区由三核苷酸3'-GAA组成。PF3基因以半保守的10核苷酸基因起始序列开始，以半保守的12核苷酸基因末端序列结束。M基因以异常的基因末端序列结束；对细胞内mRNA的分析表明，这种异常序列与通读mRNA的不成比例的高积累相关。这些研究表明，PF3编码六种独特的mRNA（NP、P+C、M、F、HN和L），它们编码七种蛋白质（NP、P、C、M、F、HN和L），并提供了PF3与仙台（鼠副流感1型）病毒密切关系的证据。
