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首页> 外文期刊>The FEBS journal >Dissecting the role of protein-protein and protein-nucleic acid interactions in MS2 bacteriophage stability
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Dissecting the role of protein-protein and protein-nucleic acid interactions in MS2 bacteriophage stability

机译：剖析蛋白质-蛋白质和蛋白质-核酸相互作用在MS2噬菌体稳定性中的作用

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摘要

To investigate the role of protein-protein and protein-nucleic acid interactions in virus assembly, we compared the stabilities of native bacteriophage MS2, virus-like particles (VLPs) containing nonviral RNAs, and an assembly-defective coat protein mutant (dlFG) and its single-chain variant (sc-dlFG). Physical (high pressure) and chemical (urea and guanidine hydrochloride) agents were used to promote virus disassembly and protein denaturation, and the changes in virus and protein structure were monitored by measuring tryptophan intrinsic fluorescence, bis-ANS probe fluorescence, and light scattering. We found that VLPs dissociate into capsid proteins that remain folded and more stable than the proteins dissociated from authentic particles. The proposed model is that the capsid disassembles but the protein remains bound to the heterologous RNA encased by VLPs. The dlFG dimerizes correctly.. but fails to assemble into capsids, because it lacks the 15-amino acid FG loop involved in inter-dimer interactions at the viral fivefold and quasi-sixfold axes. This protein was very unstable and, when compared with the dissociation/denaturation of the VLPs and the wild-type virus, it was much more susceptible to chemical and physical perturbation. Genetic fusion of the two subunits of the dimer in the single-chain dinner sc-dlFG stabilized the protein, as did the presence of 34-bp poly(GC) DNA. These Studies reveal mechanisms by which interactions in the capsid lattice can be sufficiently stable and specific to ensure assembly, and they shed light on the processes that lead to the formation of infectious viral particles.
机译：为了研究蛋白质-蛋白质和蛋白质-核酸相互作用在病毒装配中的作用，我们比较了天然噬​​菌体MS2，包含非病毒RNA的病毒样颗粒（VLP）以及装配缺陷型外壳蛋白突变体（dlFG）和它的单链变体（sc-dlFG）。使用物理（高压）和化学（尿素和胍盐酸盐）试剂来促进病毒分解和蛋白质变性，并通过测量色氨酸固有荧光，bis-ANS探针荧光和光散射来监测病毒和蛋白质结构的变化。我们发现VLPs解离成衣壳蛋白，该蛋白保持折叠状态，比从真实颗粒解离的蛋白更稳定。提出的模型是衣壳可拆卸，但蛋白质仍与VLP包裹的异源RNA结合。 dlFG正确地二聚化..但是不能组装成衣壳，因为它缺少在病毒五倍轴和准六倍轴上参与二聚体相互作用的15个氨基酸的FG环。该蛋白质非常不稳定，与VLP和野生型病毒的解离/变性相比，它更容易受到化学和物理干扰的影响。单链晚餐sc-dlFG中二聚体两个亚基的遗传融合使蛋白质稳定，34-bp poly（GC）DNA也存在。这些研究揭示了衣壳晶格中的相互作用可以足够稳定和特异以确保组装的机制，并且它们揭示了导致感染性病毒颗粒形成的过程。

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