与极其罕见的基因表达错误相关的病毒传播:单分子显微镜演示。

IF 3.8 2区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Raquel Luzón-Hidalgo, Gianluca D'Agostino, Valeria A Risso, Asuncion Delgado, Beatriz Ibarra-Molero, Luis A Campos, Jose Requejo-Isidro, Jose M Sanchez-Ruiz
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引用次数: 0

摘要

许多病毒利用程序化的移帧和停止密码子误读来合成高水平的功能性蛋白质。潜在的机制涉及复杂的RNA序列/结构基序,可能反映了低效、非编程过程的自然选择所驱动的优化。那么,根据基本的进化理论,通过基因表达错误产生的低水平蛋白质可以为病毒提供一些生存优势。在这里,我们设计了一个实验来证明这种可能性。噬菌体T7招募宿主硫氧还蛋白作为病毒DNA聚合酶的重要加工因子。我们在硫氧还蛋白基因中插入早期终止密码子,并在其末端附加编码光转换荧光蛋白的序列。病毒复制未被取消。单分子定位显微镜显示,即使每个宿主细胞平均只有大约10个硫氧还蛋白分子,噬菌体也会复制,这比典型的细胞蛋白质水平低几个数量级。我们表明,这个看似令人震惊的结果可以从分子和进化的角度来理解,因为聚合酶-硫氧还蛋白复合物表现出高动力学稳定性和长停留时间,因为这些是确保高聚合酶加工性所必需的。更一般地说,我们证明病毒复制可能是由极低拷贝数的蛋白质激活的,这表明病毒可以通过基因表达错误获得多种多样的蛋白质变体,这些蛋白质变体含有表型突变。例如,这种机制可以在跨物种传播中发挥作用,使病毒在新宿主中存活,然后在遗传水平上出现适应性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Virus Propagation Linked to Exceedingly Rare Gene-Expression Errors: A Single-Molecule Microscopy Demonstration.

Many viruses use programmed frameshifting and stop-codon misreading to synthesize functional proteins at high levels. The underlying mechanisms involve complex RNA sequence/structure motifs and likely reflect optimization driven by natural selection of inefficient, nonprogrammed processes. Then, it follows from basic evolutionary theory that low levels of proteins generated through gene expression errors could provide viruses with some survival advantage. Here, we devise an experimental demonstration of this possibility. Phage T7 recruits the host thioredoxin as an essential processivity factor for the viral DNA polymerase. We inserted early stop codons in the thioredoxin gene and appended to its end the sequence encoding for a photoconvertible fluorescent protein. Virus replication was not abolished. Single-molecule localization microscopy showed that the phage replicates even when there are only about 10 thioredoxin molecules per host cell on average, a number orders of magnitude below typical cellular protein levels. We show that this seemingly shocking result can be understood in molecular and evolutionary terms as a consequence of the polymerase-thioredoxin complex displaying high kinetic stability and a long residence time, as these are required to ensure high polymerase processivity. More generally, our demonstration that virus replication may be enabled by proteins at exceedingly low copy number suggests that viruses have access to the wide diversity of protein variants harboring phenotypic mutations as a result of gene expression errors. This mechanism could play a role, for instance, in cross-species transmission by enabling virus survival in the new host before adaptations appear at the genetic level.

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来源期刊
ACS Chemical Biology
ACS Chemical Biology 生物-生化与分子生物学
CiteScore
7.50
自引率
5.00%
发文量
353
审稿时长
3.3 months
期刊介绍: ACS Chemical Biology provides an international forum for the rapid communication of research that broadly embraces the interface between chemistry and biology. The journal also serves as a forum to facilitate the communication between biologists and chemists that will translate into new research opportunities and discoveries. Results will be published in which molecular reasoning has been used to probe questions through in vitro investigations, cell biological methods, or organismic studies. We welcome mechanistic studies on proteins, nucleic acids, sugars, lipids, and nonbiological polymers. The journal serves a large scientific community, exploring cellular function from both chemical and biological perspectives. It is understood that submitted work is based upon original results and has not been published previously.
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