Requirement of a 5'-proximal linear sequence on minus strands for plus-strand synthesis of a satellite RNA associated with turnip crinkle virus.
Requirement of a 5'-proximal linear sequence on minus strands for plus-strand synthesis of a satellite RNA associated with turnip crinkle virus.
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与萝卜皱纹病毒相关的卫星RNA的正链合成需要负链上的5-近端线性序列。
作者：
H. Guan;C. D. Carpenter;A. Simon
病毒 RNA 复制始于病毒 RNA 依赖性 RNA 聚合酶 (RdRp) 和/或相关宿主因子对顺式作用 RNA 元件的特异性识别。一个短 RNA 元件 (3'-AACCCCUGGGAGGC) 位于距卫星 RNA C (satC) 负链 5' 端 41 个碱基处，卫星 RNA C 是一种与芜菁皱病毒 (TCV) 天然相关的 356 个碱基的亚病毒 RNA，以前使用体外 RdRp 测定法鉴定出对正链合成很重要（H.guan、C.Song、A.E.Simon，1997，RNA 3， 1401-1412）。为了检查该元件在 RNA 复制中的功能意义，将突变引入该元件的连续 C 残基中。当在原生质体中检测转录物并抑制体外该元件指导的转录时，3'端 C 残基的单一突变导致 satC + 链水平不可检测。然而，在原生质体接种后 6 小时 (hpi) 时可检测到 satC 负链，在 24 hpi 时累积至野生型水平的约 10%。当处于正义方向时，这种突变对体外全长 satC 正链的负链合成影响很小或没有影响，表明 5'-近端 RNA 元件是 satC 正链合成所必需的。此外，体内遗传选择揭示了对该元件 14 个核苷酸中的 10 个的严格要求，表明一级序列对于 RNA 积累至关重要。
Viral RNA replication begins with specific recognition of cis-acting RNA elements by the viral RNA-dependent RNA polymerase (RdRp) and/or associated host factors. A short RNA element (3'-AACCCCUGGGAGGC) located 41 bases from the 5' end of minus strands of satellite RNA C (satC), a 356-base subviral RNA naturally associated with turnip crinkle virus (TCV), was previously identified as important for plus-strand synthesis using an in vitro RdRp assay (H. Guan, C. Song, A. E. Simon, 1997, RNA 3, 1401-1412). To examine the functional significance of this element in RNA replication, mutations were introduced into the consecutive C residues in the element. A single mutation of the 3'-most C residue resulted in undetectable levels of satC plus strands when transcripts were assayed in protoplasts and suppressed transcription directed by the element in vitro. However, satC minus strands were detectable at 6 h postinoculation (hpi) of protoplasts, accumulating to about 10% of wild-type levels at 24 hpi. This mutation, when in the plus-sense orientation, had little or no effect on minus-strand synthesis from full-length satC plus strands in vitro, suggesting that the 5'-proximal RNA element is required for satC plus-strand synthesis. In addition, in vivo genetic selection revealed a strict requirement for 10 of the 14 nucleotides of the element, indicating that the primary sequence is essential for RNA accumulation.
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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
发表时间：
1997
期刊：
Virology.
影响因子：
--
作者：
Singh,RN;Dreher,TW
通讯作者：
Dreher,TW
由非特异性二级结构和-CCR-盒组合产生的RNA中的特异性位点选择：萝卜黄花叶病毒RNA依赖性RNA聚合酶启动负链合成。
DOI：
10.1017/s1355838298980694
发表时间：
1998
期刊：
RNA (New York, N.Y.)
影响因子：
--
作者：
Singh,RN;Dreher,TW
通讯作者：
Dreher,TW
BMV RNA3 亚基因组启动子的核心和调节序列的突变分析。
影响因子：
14.9
作者：
Marsh,LE;Dreher,TW;Hall,TC
通讯作者：
Hall,TC
影响因子：
2.9
作者：
BROWN, D;GOLD, L
通讯作者：
GOLD, L
影响因子：
14.9
作者：
ZUKER, M;JAEGER, JA;TURNER, DH
通讯作者：
TURNER, DH