Takahashi T, Oie M, Ichihashi Y
Central Research Laboratory, Takasago International Company, Tokyo, Japan.
Virology. 1994 Aug 1;202(2):844-52. doi: 10.1006/viro.1994.1406.
The N-terminal amino acid sequences of vaccinia virus structural proteins were determined by direct sequencing following separation of the proteins of purified intracellular mature virus by SDS-polyacrylamide gels. By comparing the sequences obtained with the published vaccinia virus DNA sequences, specific open reading frames (ORFs) were identified. The structural proteins were encoded by the ORFs of HindIII, A3L (VP57K, 32K), A10L (VP62K, VP28K, VP22K), A12L (VP10K, VP4K), A13L (VP14K), A14L (VP17-25K), A17L (VP23-29K), A27L (VP13.8K), D8L (VP32K), H3L (VP34-37K), L4R (VP27K), G7L (VP16K), and 15L (VP13K). Four virus membrane proteins contained transmembrane signals. The N-termini of proteins indicated four types of cleavages. Ala-Gly-specific cleavage associated with products of six ORFs. Phe-specific cleavage was found in two, Met-specific in three, and Arg-specific in the product of one ORF. Ala-Gly-specific cleavage processes seven core proteins encoded by five ORFs and one membrane protein. The Met- and Arg-specific cleavages are suggested to be nonessential for virus assembly because the major portions of the target membrane proteins remain unaffected.
通过十二烷基硫酸钠-聚丙烯酰胺凝胶对纯化的细胞内成熟病毒的蛋白质进行分离后，直接测序确定了痘苗病毒结构蛋白的N端氨基酸序列。通过将获得的序列与已发表的痘苗病毒DNA序列进行比较，确定了特定的开放阅读框（ORF）。结构蛋白由HindIII、A3L（VP57K、32K）、A10L（VP62K、VP28K、VP22K）、A12L（VP10K、VP4K）、A13L（VP14K）、A14L（VP17 - 25K）、A17L（VP23 - 29K）、A27L（VP13.8K）、D8L（VP32K）、H3L（VP34 - 37K）、L4R（VP27K）、G7L（VP16K）和15L（VP13K）的ORF编码。四种病毒膜蛋白含有跨膜信号。蛋白质的N端显示出四种类型的切割。六种ORF的产物存在丙氨酸-甘氨酸特异性切割。在两种产物中发现苯丙氨酸特异性切割，在三种产物中发现甲硫氨酸特异性切割，在一种ORF的产物中发现精氨酸特异性切割。丙氨酸-甘氨酸特异性切割处理由五个ORF和一种膜蛋白编码的七种核心蛋白。甲硫氨酸和精氨酸特异性切割被认为对病毒组装不是必需的，因为目标膜蛋白的主要部分未受影响。
