The Effects of Packaged, but Misguided, Single-Stranded DNA Genomes Are Transmitted to the Outer Surface of the ϕX174 Capsid
The Effects of Packaged, but Misguided, Single-Stranded DNA Genomes Are Transmitted to the Outer Surface of the ϕX174 Capsid
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包装但被误导的单链 DNA 基因组的影响被传输到 X174 衣壳的外表面
DOI：
10.1128/jvi.00883-21
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发表时间：
2021
影响因子：
5.4
通讯作者：
Fane, Bentley A.
中科院分区：
文献类型：
--
作者：
Ogunbunmi, Elizabeth T.;Roznowski, Aaron P.;Fane, Bentley A.
大多数二十面体病毒将其基因组浓缩成体积受限的衣壳。然而，同时发生的基因组生物合成和包装是单链DNA（ssDNA）病毒特有的。ssDNA基因组包装结合了双链DNA（dsDNA）和ssRNA系统中发现的元件。与dsDNA病毒类似，基因组被包装到预先形成的衣壳中。像ssRNA病毒一样，有许多衣壳-基因组关联。在ssDNA微病毒中，DNA结合蛋白J在60个二十面体有序的DNA结合口袋之间引导基因组。它还部分中和DNA的负磷酸骨架。与X174相关的微病毒，如G4和α3，具有长度和电荷结构不同的J蛋白。这表明互换J蛋白可以改变用于引导衣壳中DNA的路径。先前，表征了其中G4基因替换了G4 X174 J基因的G4 X4 J嵌合体。它显示出致命的包装缺陷，导致原壳被从生产组装中移除。在这里，我们报告了另一种不能存活的嵌合体，α 3 J。与α XG 4J不同，α XG 3 J能有效地包装DNA，但产生非感染性颗粒。这些颗粒显示出附着宿主细胞的能力降低，表明内部DNA组织可能会扭曲衣壳的外表面。恢复活力的突变改变了J-外壳蛋白接触位点。这些结果为ssDNA的组织可以影响包装和包装后的现象提供了证据。重要的ssDNA病毒利用二十面体有序的蛋白质-核酸相互作用来引导和组织它们的基因组进入预先形成的外壳。如前所述，混乱的基因组衣壳协会可以通过破坏包装复合物的稳定来抑制BMPX 174包装。然而，组织不良的基因组的后果可能超出包装反应。如本文所证明的，其可导致非感染性包装颗粒。因此，ssDNA基因组应该被认为是一个完整的和结构性的病毒体组分，影响整个颗粒的性质，包括衣壳的外表面。
Most icosahedral viruses condense their genomes into volumetrically constrained capsids. However, concurrent genome biosynthesis and packaging are specific to single-stranded DNA (ssDNA) viruses. ssDNA genome packaging combines elements found in both double-stranded DNA (dsDNA) and ssRNA systems. Similar to dsDNA viruses, the genome is packaged into a preformed capsid. Like ssRNA viruses, there are numerous capsid-genome associations. In ssDNA microviruses, the DNA-binding protein J guides the genome between 60 icosahedrally ordered DNA binding pockets. It also partially neutralizes the DNA’s negative phosphate backbone. ϕX174-related microviruses, such as G4 and α3, have J proteins that differ in length and charge organization. This suggests that interchanging J proteins could alter the path used to guide DNA in the capsid. Previously, a ϕXG4J chimera, in which the ϕX174 J gene was replaced with the G4 gene, was characterized. It displayed lethal packaging defects, which resulted in procapsids being removed from productive assembly. Here, we report the characterization of another inviable chimera, ϕXα3J. Unlike ϕXG4J, ϕXα3J efficiently packaged DNA but produced noninfectious particles. These particles displayed a reduced ability to attach to host cells, suggesting that internal DNA organization could distort the capsid’s outer surface. Mutations that restored viability altered J-coat protein contact sites. These results provide evidence that the organization of ssDNA can affect both packaging and postpackaging phenomena.IMPORTANCEssDNA viruses utilize icosahedrally ordered protein-nucleic acids interactions to guide and organize their genomes into preformed shells. As previously demonstrated, chaotic genome-capsid associations can inhibit ϕX174 packaging by destabilizing packaging complexes. However, the consequences of poorly organized genomes may extend beyond the packaging reaction. As demonstrated herein, it can lead to uninfectious packaged particles. Thus, ssDNA genomes should be considered an integral and structural virion component, affecting the properties of the entire particle, which includes the capsid’s outer surface.
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DOI：
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DOI：
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发表时间：
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