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利用病毒基因的cDNA克隆及融合蛋白生物学重要区域的序列对仙台病毒mRNA进行分析。

Analysis of Sendai virus mRNAs with cDNA clones of viral genes and sequences of biologically important regions of the fusion protein.

作者信息

Hsu M, Choppin P W

出版信息

Proc Natl Acad Sci U S A. 1984 Dec;81(24):7732-6. doi: 10.1073/pnas.81.24.7732.

Abstract

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.

摘要

分离出代表仙台病毒五个基因(P、HN、NP、F和M)的cDNA克隆,并用于通过杂交鉴定病毒mRNA。鉴定出了这五个基因的单顺反子转录本的五种mRNA。基于大小以及与病毒RNA而非其他五个基因的cDNA的杂交鉴定出的第六种转录本,被认为代表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。给出了F蛋白两个生物学重要区域的核苷酸序列和推导的氨基酸序列——F蛋白COOH末端近端约25%的区域以及跨越激活该蛋白融合活性的蛋白水解切割位点的区域。F蛋白在其通过疏水区域锚定在病毒膜中的部分之外,有一个由42个氨基酸组成的异常大的“细胞质结构域”。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26个疏水氨基酸从该切割位点在F1多肽的NH2末端延伸。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ab76/392226/f8accf7968f1/pnas00625-0072-a.jpg

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