Insights into polyprotein processing and RNA-protein interactions in foot-and-mouth disease virus genome replication
Insights into polyprotein processing and RNA-protein interactions in foot-and-mouth disease virus genome replication
复制标题
深入了解口蹄疫病毒基因组复制中的多蛋白加工和 RNA-蛋白质相互作用
DOI：
10.1101/2023.01.26.525530
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Pierce D
中科院分区：
文献类型：
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作者：
Pierce D
口蹄疫病毒（Foot-and-mouth disease virus，FMDV）是一种小核糖核酸病毒，感染偶蹄类动物引起口蹄疫。正义RNA基因组包含单个开放阅读框，其被翻译为多蛋白，该多蛋白被病毒蛋白酶切割以产生病毒结构和非结构蛋白。最初的加工发生在三个主要的交界处，产生四个主要的前体; Lpro和P1，P2和P3（也称为1ABCD，2BC和3AB 1，2，3CD）。2BC和3AB 1，2，3CD前体随后进行蛋白水解以产生病毒复制所需的蛋白质，包括酶2C，3Cpro和3Dpol。这些前体可以通过顺式和反式（即，分子内和分子间蛋白水解）途径，其被认为对于控制病毒复制是重要的。我们以前的研究表明，3B 3 -3C连接中的单个残基在控制3AB 1，2，3CD加工中具有重要作用。在这里，我们使用基于体外的测定来表明，在3B 3 -3C边界处的单个氨基酸取代增加了蛋白水解的速率，以产生一种新的含2C的前体。互补分析表明，虽然这种氨基酸取代增强了一些非酶的非结构蛋白的生产，那些酶的功能被抑制。有趣的是，复制只能通过与切割RNA元件的突变互补来支持，这为复制酶和RNA元件之间的功能相互作用提供了遗传学证据。重要信息口蹄疫病毒（Foot-and-mouth disease virus，FMDV）是引起口蹄疫（Foot-and-mouth disease，FMD）的重要疾病，该疾病在世界许多地区流行，可导致重大经济损失。病毒的复制发生在受感染细胞的膜相关区室中，需要高度协调的加工事件来产生一系列非结构蛋白。这些最初作为多蛋白产生，其可能通过顺式和反式途径进行蛋白水解（即，分子内和分子间蛋白水解）。替代加工途径的作用可能有助于通过提供蛋白质生产的时间控制来协调病毒复制，在这里，我们分析了改变FMDV中这些途径的氨基酸取代的后果。我们的数据表明，正确的处理是需要产生关键酶复制的环境中，他们可以相互作用的基本病毒RNA元素。这些数据进一步了解RNA基因组复制。
Foot-and-mouth disease virus (FMDV) is a picornavirus, which infects cloven-hoofed animals to cause foot-and-mouth disease (FMD). The positive-sense RNA genome contains a single open reading frame, which is translated as a polyprotein that is cleaved by viral proteases to produce the viral structural and nonstructural proteins. Initial processing occurs at three main junctions to generate four primary precursors; Lproand P1, P2, and P3 (also termed 1ABCD, 2BC, and 3AB1,2,3CD). The 2BC and 3AB1,2,3CD precursors undergo subsequent proteolysis to generate the proteins required for viral replication, including the enzymes 2C, 3Cpro, and 3Dpol. These precursors can be processed through bothcisandtrans(i.e., intra- and intermolecular proteolysis) pathways, which are thought to be important for controlling virus replication. Our previous studies suggested that a single residue in the 3B3-3C junction has an important role in controlling 3AB1,2,3CD processing. Here, we usein vitrobased assays to show that a single amino acid substitution at the 3B3-3C boundary increases the rate of proteolysis to generate a novel 2C-containing precursor. Complementation assays showed that while this amino acid substitution enhanced production of some nonenzymatic nonstructural proteins, those with enzymatic functions were inhibited. Interestingly, replication could only be supported by complementation with mutations incisacting RNA elements, providing genetic evidence for a functional interaction between replication enzymes and RNA elements.IMPORTANCEFoot-and-mouth disease virus (FMDV) is responsible for foot-and-mouth disease (FMD), an important disease of farmed animals, which is endemic in many parts of the world and can results in major economic losses. Replication of the virus occurs within membrane-associated compartments in infected cells and requires highly coordinated processing events to produce an array of nonstructural proteins. These are initially produced as a polyprotein that undergoes proteolysis likely through bothcisandtransalternative pathways (i.e., intra- and intermolecular proteolysis). The role of alternative processing pathways may help coordination of viral replication by providing temporal control of protein production and here we analyze the consequences of amino acid substitutions that change these pathways in FMDV. Our data suggest that correct processing is required to produce key enzymes for replication in an environment in which they can interact with essential viral RNA elements. These data further the understanding of RNA genome replication.