Enami K, Sato T A, Nakada S, Enami M
Department of Biochemistry, Kanazawa University School of Medicine, Ishikawa, Japan.
J Virol. 1994 Mar;68(3):1432-7. doi: 10.1128/JVI.68.3.1432-1437.1994.
The influenza virus NS1 protein was shown to stimulate translation of the M1 protein. M-CAT RNA, which contains the chloramphenicol acetyltransferase (CAT) reporter gene and the terminal noncoding sequence of segment 7 (coding for the M1 and M2 proteins), was ribonucleoprotein transfected into clone 76 cells expressing the influenza virus RNA polymerase and NP proteins required for the transcription and replication of influenza virus ribonucleoproteins. When the cells were superinfected with a recombinant vaccinia virus which expresses the NS1 protein, CAT expression from the M-CAT RNA was significantly stimulated but transcription was not altered. The expression of NS-CAT RNA, which contains noncoding sequences of segment 8 (coding for the NS1 and NS2 proteins), was not altered by the NS1 protein. Site-directed mutagenesis showed that the sequence GGUAGAUA upstream of the initiation codon on segment 7 was required for stimulation.
流感病毒NS1蛋白被证明可刺激M1蛋白的翻译。将含有氯霉素乙酰转移酶(CAT)报告基因和第7节段末端非编码序列(编码M1和M2蛋白)的M-CAT RNA核糖核蛋白转染到表达流感病毒核糖核蛋白转录和复制所需的流感病毒RNA聚合酶和NP蛋白的76克隆细胞中。当用表达NS1蛋白的重组痘苗病毒对细胞进行超感染时,M-CAT RNA的CAT表达受到显著刺激,但转录未改变。含有第8节段非编码序列(编码NS1和NS2蛋白)的NS-CAT RNA的表达不受NS1蛋白的影响。定点诱变表明,第7节段起始密码子上游的GGUAGAUA序列是刺激所必需的。