Analysis of Sendai virus mRNAs with cDNA clones of viral genes and sequences of biologically important regions of the fusion protein.
Analysis of Sendai virus mRNAs with cDNA clones of viral genes and sequences of biologically important regions of the fusion protein.
复制标题
使用病毒基因的 cDNA 克隆和融合蛋白的生物学重要区域的序列分析仙台病毒 mRNA。
DOI：
10.1073/pnas.81.24.7732
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发表时间：
1984
影响因子：
11.1
通讯作者：
Choppin,PW
中科院分区：
文献类型：
--
作者：
Hsu,M;Choppin,PW
分离代表仙台病毒的五个基因（P、HN、NP、F和M）的cDNA克隆，并用于通过杂交鉴定病毒mRNA。鉴定了这些基因的单顺反子转录物的五种mRNA。第六个转录本是根据大小和与病毒RNA杂交而不是与其他五个基因的cDNA杂交而鉴定的，被认为代表了L蛋白的信息。此外，还发现了NP和P基因以及M和F基因的多顺反子转录本，后者确定了F基因与M基因相邻的位置，这些结果证实并扩展了先前报道的病毒部分基因顺序。核苷酸序列和衍生的氨基酸序列的两个生物学上重要的区域的F蛋白-约25%的F近端其COOH末端和跨越该位点的蛋白水解裂解，激活蛋白质的融合活性的区域-提出。F蛋白在疏水区之外有一个非常大的42个氨基酸的“胞质结构域”，它通过疏水区锚定在病毒膜上。在F1和F2多肽的连接处发现了一个可能的胰蛋白酶切割位点，并且在F1多肽的NH 2末端从该切割位点延伸出26个疏水性氨基酸。
cDNA clones representing five of the genes of Sendai virus (P, HN, NP, F, and M) were isolated and used to identify the viral mRNAs by hybridization. Five mRNAs that were monocistronic transcripts of these genes were identified. A sixth transcript, which was identified on the basis of size and of hybridization to viral RNA but not to the cDNA of the other five genes, is thought to represent the message for the L protein. In addition, polycistronic transcripts of the NP and P genes and of the M and F genes were also found. The latter establishes the position of the F gene adjacent to the M gene; these results confirm and extend the previously reported partial gene order of the virus. Nucleotide sequences and derived amino acid sequences of two biologically important regions of the F protein--approximately 25% of F proximal to its COOH terminus and the region spanning the site of the proteolytic cleavage that activates the fusion activity of the protein--are presented. The F protein has an unusually large "cytoplasmic domain" of 42 amino acids beyond the hydrophobic region by which it is anchored in the viral membrane. A single possible trypsin cleavage site was found at the junction of the F1 and F2 polypeptides, and 26 hydrophobic amino acids extend from this cleavage site at the NH2 terminus of the F1 polypeptide.