Kadaré G, David C, Haenni A L
Institut Jacques Monod, Paris, France.
J Virol. 1996 Nov;70(11):8169-74. doi: 10.1128/JVI.70.11.8169-8174.1996.
The 206-kDa protein of turnip yellow mosaic virus belongs to an expanding group of proteins containing a domain which includes the consensus nucleotide binding site GxxxxGKS/T. A portion of this protein (amino acids [aa] 916 to 1259) was expressed in Escherichia coli and purified by affinity chromatography to near homogeneity. In the absence of any other viral factors, it exhibited ATPase and GTPase activities in vitro. A mutant protein with a single amino acid substitution in the consensus nucleotide binding site (Lys-982 to Ser) exhibited only low levels of both activities, implying that Lys-982 is important for nucleoside triphosphatase activity. The protein also possessed nonspecific RNA binding capacity. Deletion mutants revealed that an N-terminal domain (aa 916 to 1061) and a C-terminal domain (aa 1182 to 1259) participate in RNA binding. The results presented here provide the first experimental evidence that turnip yellow mosaic virus encodes nucleoside triphosphatase and RNA binding activities.
芜菁黄花叶病毒的206 kDa蛋白属于一个不断扩大的蛋白家族,该家族蛋白含有一个结构域,其中包括保守的核苷酸结合位点GxxxxGKS/T。该蛋白的一部分(氨基酸[aa] 916至1259)在大肠杆菌中表达,并通过亲和层析纯化至近乎纯品。在没有任何其他病毒因子的情况下,它在体外表现出ATP酶和GTP酶活性。在保守核苷酸结合位点发生单个氨基酸取代的突变蛋白(Lys-982突变为Ser)仅表现出低水平的这两种活性,这意味着Lys-982对核苷三磷酸酶活性很重要。该蛋白还具有非特异性RNA结合能力。缺失突变体表明,N端结构域(aa 916至1061)和C端结构域(aa 1182至1259)参与RNA结合。此处给出的结果提供了首个实验证据,证明芜菁黄花叶病毒编码核苷三磷酸酶和RNA结合活性。