Isolation from Tobacco Mosaic Virus-Infected Tobacco of a Solubilized Template-Specific RNA-Dependent RNA Polymerase Containing a 126K/183K Protein Heterodimer
Isolation from Tobacco Mosaic Virus-Infected Tobacco of a Solubilized Template-Specific RNA-Dependent RNA Polymerase Containing a 126K/183K Protein Heterodimer
复制标题
从感染烟草花叶病毒的烟草中分离出含有 126K/183K 蛋白异二聚体的可溶模板特异性 RNA 依赖性 RNA 聚合酶
DOI：
10.1128/jvi.73.4.2633-2640.1999
复制
发表时间：
1999
影响因子：
5.4
通讯作者：
A. Ishihama
中科院分区：
文献类型：
--
作者：
Takato Watanabe;A. Honda;A. Iwata;S. Ueda;T. Hibi;A. Ishihama
摘要 确定了烟草花叶病毒（TMV）OM株推定RNA聚合酶（183K蛋白）基因的完整核苷酸序列，该菌株与相关菌株vulgare的核苷酸序列有51个位置不同，氨基酸序列有6个残基。该 183K 蛋白的三个片段，每个片段均包含甲基转移酶 (M)、解旋酶 (H) 或 RNA 依赖性 RNA 聚合酶 (P) 的序列基序，在大肠杆菌中表达为带有六组氨酸标签的融合蛋白，并针对纯化的带有组氨酸标签的 M 和 P 多肽产生了域特异性抗体。通过免疫亲和纯化，分离出包含全长183K和126K（183K蛋白的氨基末端近端部分）病毒蛋白的异二聚体的模板特异性RNA依赖性RNA聚合酶。我们认为，用于合成负链 RNA 的 TMV RNA 聚合酶由 183 kDa 和 126 kDa 蛋白各一个分子组成，可能与两个或多个宿主蛋白一起组成。
ABSTRACT The complete nucleotide sequence was determined for the putative RNA polymerase (183K protein) gene of tobacco mosaic virus (TMV) OM strain, which differed from the related strain, vulgare, by 51 positions in its nucleotide sequence and 6 residues in its amino acid sequence. Three segments of this 183K protein, each containing the sequence motif of methyltransferase (M), helicase (H), or RNA-dependent RNA polymerase (P), were expressed in Escherichia coli as fusion proteins with hexahistidine tags, and domain-specific antibodies were raised against purified His-tagged M and P polypeptides. By immunoaffinity purification, a template-specific RNA-dependent RNA polymerase containing a heterodimer of the full-length 183K and 126K (an amino-terminal-proximal portion of the 183K protein) viral proteins was isolated. We propose that the TMV RNA polymerase for minus-strand RNA synthesis is composed of one molecule each of the 183- and 126-kDa proteins, possibly together with two or more host proteins.
DOI：
10.1073/pnas.90.4.1498
发表时间：
1993-02-15
影响因子：
11.1
作者：
QUADT, R;KAO, CC;AHLQUIST, P
通讯作者：
AHLQUIST, P