Spriggs M K, Olmsted R A, Venkatesan S, Coligan J E, Collins P L
Virology. 1986 Jul 15;152(1):241-51. doi: 10.1016/0042-6822(86)90388-0.
The complete sequences of the fusion (F) mRNA and protein of human parainfluenza virus type 3 (PF3) were determined from overlapping cDNA clones. To confirm the cDNA sequence, the complete sequence of the F gene was determined independently by dideoxynucleotide sequencing of genomic RNA using synthetic oligonucleotide primers. The mRNA contains 1845 nucleotides, exclusive of poly (A), has an unusually long (193-nucleotide) 5' nontranslated region, and encodes an F0 protein of 539 amino acids. The site within F0 of the proteolytic cleavage that activates fusion activity was established by direct amino acid sequencing of the NH2 terminus of the F1 subunit. The PF3 F0 protein shares major structural features with the previously sequenced F0 proteins of Sendai virus (murine parainfluenza type 1) and simian virus 5 (SV5, canine parainfluenza type 2), including: similarity in overall length; similarity in location of the site of the activating proteolytic cleavage; the presence of an NH2-terminal signal peptide and COOH-proximal membrane anchor; strong conservation of the sequence at the NH2 terminus of the F1 subunit; and nearly exact conservation in the number and positions of cysteine residues. Alignment of the F0 protein sequences of PF3 with those of Sendai, SV5, and respiratory syncytial virus (RSV) using a matrix that scores both amino acid matches and mismatches provided highly significant statistical evidence that all four proteins are related. The order of decreasing relatedness to PF3 was found to be: Sendai virus, SV5, and RSV.
通过重叠cDNA克隆确定了人副流感病毒3型(PF3)融合(F)mRNA和蛋白的完整序列。为了确认cDNA序列,使用合成寡核苷酸引物通过对基因组RNA进行双脱氧核苷酸测序独立确定了F基因的完整序列。该mRNA含有1845个核苷酸(不包括聚腺苷酸),具有异常长的(193个核苷酸)5'非翻译区,并编码一个由539个氨基酸组成的F0蛋白。通过对F1亚基NH2末端进行直接氨基酸测序确定了激活融合活性的蛋白水解切割在F0内的位点。PF3 F0蛋白与先前测序的仙台病毒(鼠副流感1型)和猿猴病毒5(SV5,犬副流感2型)的F0蛋白具有主要结构特征,包括:全长相似性;激活蛋白水解切割位点位置的相似性;存在NH2末端信号肽和COOH近端膜锚;F1亚基NH2末端序列的高度保守;以及半胱氨酸残基数量和位置几乎完全保守。使用对氨基酸匹配和错配都计分的矩阵将PF3的F0蛋白序列与仙台病毒、SV5和呼吸道合胞病毒(RSV)的序列进行比对,提供了高度显著的统计证据表明这四种蛋白是相关的。发现与PF3相关性从高到低的顺序为:仙台病毒、SV5和RSV。