Multiple Cis-acting elements modulate programmed -1 ribosomal frameshifting in Pea enation mosaic virus.
Multiple Cis-acting elements modulate programmed -1 ribosomal frameshifting in Pea enation mosaic virus.
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DOI：
10.1093/nar/gkv1241
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发表时间：
2016-01-29
影响因子：
14.9
通讯作者：
Simon AE
中科院分区：
文献类型：
--
作者：
Gao F;Simon AE
程序性-1核糖体移码（-1 PRF）被许多正链RNA病毒用于翻译所需产物。尽管进行了广泛的研究，但仍然没有解决编码位点下游的顺式元件如何促进精确水平的移码。豌豆冠状花叶病毒RNA 2通过5′端ORF（p33）的-1PRF表达其RNA聚合酶。位于重新编码位点附近的三个发夹结构在影病毒中是遗传保守的。位于p33终止密码子下游的中央转录刺激元件（RSE）是具有两个不对称内环的大发夹。突变分析表明，整个RSE和RSE下茎（LS）结构的序列是重要的移码。突变体的SHAPE探测表明存在更高级的结构，并且LS中的序列也可以适应另一种构象。当LS结构稳定时，RSE和3′末端发夹之间的长距离配对不那么重要。在不存在RSE的情况下发生的基础水平的移码增加到野生型的72%时，也删除了发夹上游的滑网站。这些结果表明，在没有活性RSE构象的情况下，可能需要抑制移码。
Programmed -1 ribosomal frameshifting (-1 PRF) is used by many positive-strand RNA viruses for translation of required products. Despite extensive studies, it remains unresolved how cis-elements just downstream of the recoding site promote a precise level of frameshifting. The Umbravirus Pea enation mosaic virus RNA2 expresses its RNA polymerase by -1 PRF of the 5′-proximal ORF (p33). Three hairpins located in the vicinity of the recoding site are phylogenetically conserved among Umbraviruses. The central Recoding Stimulatory Element (RSE), located downstream of the p33 termination codon, is a large hairpin with two asymmetric internal loops. Mutational analyses revealed that sequences throughout the RSE and the RSE lower stem (LS) structure are important for frameshifting. SHAPE probing of mutants indicated the presence of higher order structure, and sequences in the LS may also adapt an alternative conformation. Long-distance pairing between the RSE and a 3′ terminal hairpin was less critical when the LS structure was stabilized. A basal level of frameshifting occurring in the absence of the RSE increases to 72% of wild-type when a hairpin upstream of the slippery site is also deleted. These results suggest that suppression of frameshifting may be needed in the absence of an active RSE conformation.