Modulation of replication, aminoacylation and adenylation in vitro and infectivity in vivo of BMV RNAs containing deletions within the multifunctional 3' end.
Modulation of replication, aminoacylation and adenylation in vitro and infectivity in vivo of BMV RNAs containing deletions within the multifunctional 3' end.
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调节多功能 3 末端缺失的 BMV RNA 的体外复制、氨酰化和腺苷酸化以及体内感染性。
DOI：
10.1002/j.1460-2075.1986.tb04425.x
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Kaesberg,P
中科院分区：
文献类型：
--
作者：
Bujarski,JJ;Ahlquist,P;Hall,TC;Dreher,TW;Kaesberg,P
雀麦花叶病毒（BMV）的基因组由三条（+）链RNA组成，每条RNA在其3′端含有相似的、高度结构化的、200个碱基长的序列。该序列的134个碱基亚组包含指导病毒RNA与BMV RNA复制酶、ATP、CTP：tRNA核苷酸转移酶和氨酰tRNA合成酶相互作用的信号。一系列的突变体包含在这个区域内的缺失，以前构建和测试在体外的复制和氨酰化活性的影响，现在已经在体外测定腺苷酸化功能和在体内的能力，在分离的原生质体和整个植物中复制。这些试验表明，BMV复制酶识别的病毒RNA的特征与指导腺苷酸化的特征重叠，但与指导氨酰化的特征不同。因此，优先抑制氨酰化的缺失的致死性表明该功能可能对体内病毒复制具有重要作用。携带20个碱基缺失的RNA 3突变体在体外产生正常水平的氨酰化和增强水平的复制酶模板活性和腺苷酸化，能够在原生质体和植物中复制;然而，其在原生质体中的积累相对于野生型减少。这表明影响病毒RNA复制和积累的其他功能存在于保守的3′序列中。
The genome of brome mosaic virus (BMV) is comprised of three (+) strand RNAs, each containing a similar, highly structured, 200 base long sequence at its 3′ end. A 134 base subset of this sequence contains signals directing interaction of the viral RNA with BMV RNA replicase, ATP,CTP:tRNA nucleotidyl transferase and aminoacyl tRNA synthetase. A series of mutants containing deletions within this region, previously constructed and tested in vitro for the effect on replication and aminoacylation activities, has now been assayed in vitro for adenylation function and in vivo for ability to replicate in isolated protoplasts and whole plants. These tests indicate that features of viral RNA recognized by BMV replicase overlap those directing adenylation, but are distinct from those directing aminoacylation. Consequently, the lethality of a deletion preferentially inhibiting aminoacylation suggests that this function may have an essential role contributing to viral replication in vivo. An RNA3 mutant bearing a 20‐base deletion yielding normal levels of aminoacylation and enhanced levels of replicase template activity and adenylation in vitro was able to replicate in protoplasts and plants; however, its accumulation in protoplasts was reduced relative to wild‐type. This suggests that additional functions affecting the replication and accumulation of viral RNA reside in the conserved 3′ sequence.
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