Turnip yellow mosaic virus RNA-dependent RNA polymerase: initiation of minus strand synthesis in vitro.
Turnip yellow mosaic virus RNA-dependent RNA polymerase: initiation of minus strand synthesis in vitro.
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芜菁黄花叶病毒RNA依赖性RNA聚合酶：体外负链合成的起始。
DOI：
10.1006/viro.1997.8621
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发表时间：
1997
期刊：
影响因子：
--
通讯作者：
Dreher,TW
中科院分区：
文献类型：
--
作者：
Singh,RN;Dreher,TW
研究了萝卜黄花叶病毒（TYMV）侵染大白菜叶片叶绿体膜的RNA依赖性RNA聚合酶（RdRp）活性。模板依赖性的微球菌核酸酶处理活性从TYMV RNA和短至28个核苷酸长的RNA的3 ‘片段合成全长负链，包括3 ’ - trna样结构（TLS）的氨基酸受体茎。当GTP插入到3 '端-CCA的倒数第二个(C)残基的对面时，负链被证明是由新起始产生的。TYMV RdRp活性是模板特异性的，poly(A) RNA不被复制，而AlMV (AlMV) RNA不包含3 ' -TLS，是一个非常差的模板。然而，其他具有3 ' -TLS的病毒rna和trna的体外转录本都有不同程度的复制。完全修饰的trna要么是无活性模板，要么是活性较差的模板，而AlMV 3 ‘ -RNA，即使提供了3 ’末端-ACCA，也无法被检测到复制。受体茎伪结作为启动子元件的潜在作用是通过突变来评估的，这些突变极大地改变了伪结的结构和序列，揭示了在降低模板活性方面只有双重作用。数据表明，具有trna样构象和-CCA 3 '末端的rna都是TYMV RdRp的潜在模板，并表明启动子元件并不局限于受体干假结。
An RNA-dependent RNA polymerase (RdRp) activity was detergent-solubilized from the chloroplast membranes of Chinese cabbage leaves infected with turnip yellow mosaic virus (TYMV). The template-dependent, micrococcal nuclease-treated activity synthesized full-length minus strands from TYMV RNA and 3′-fragments as short as a 28-nucleotide-long RNA comprising the amino acid acceptor stem of the 3′-tRNA-like structure (TLS). Minus strands were shown to arise byde novoinitiation with the insertion of GTP opposite the penultimate (C) residue of the 3′-terminal -CCA. The TYMV RdRp activity was template specific in that poly(A) RNA was not copied, and alfalfa mosaic virus (AlMV) RNA, which does not contain a 3′-TLS, was a very poor template. However, other viral RNAs with a 3′-TLS andin vitrotranscripts of tRNAs were copied to varying degrees. Fully modified tRNAs were either inactive or poorly active templates, and AlMV 3′-RNA, even when provided with a 3′-terminal -ACCA, was not copied detectably. A potential role of the acceptor stem pseudoknot as a promoter element was assessed with mutations that drastically altered the structure and sequence of the pseudoknot, revealing only a twofold effect in decreasing template activity. The data show that RNAs with both a tRNA-like conformation and a -CCA 3′-terminus are potential templates for TYMV RdRp and suggest that promoter elements are not limited to the acceptor stem pseudoknot.
对来自萝卜黄花叶病毒 RNA 的克隆 cDNA 的 tRNA 样转录物进行的验证表明，病毒 RNA 3 端的 L 形区域不足以实现最佳氨酰化。
影响因子：
3.9
作者：
T. Dreher;C. Florentz;R. Giegé
通讯作者：
R. Giegé
使用微球菌核酸酶从受雀麦花叶病毒感染的大麦中纯化高度模板依赖的 RNA 依赖性 RNA 聚合酶。
影响因子：
3.7
作者：
W. Miller;T. Hall
通讯作者：
T. Hall
萝卜黄花叶病毒RNA tRNA样结构中假结的突变分析。
影响因子：
5.6
作者：
R. Mans;Marleen H. Van Steeg;P. Verlaan;C. Pleij;L. Bosch
通讯作者：
L. Bosch