Construction of a Fusellovirus with a Minimal Set of Genes.
Construction of a Fusellovirus with a Minimal Set of Genes.
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DOI：
10.1021/acssynbio.1c00232
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发表时间：
2021-09
影响因子：
4.7
通讯作者：
Junxia Zhang;Li Huang
中科院分区：
文献类型：
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作者：
Junxia Zhang;Li Huang
在全球各地的酸性温泉中发现了大量的聚合体病毒。它们共享一组高度保守的基因(核心基因)，并拥有不同数量的保守程度较低的基因(非核心基因)。然而，这些基因中的大多数功能尚不清楚。最近的研究表明，这些基因中有多达一半能耐受突变。在本研究中，我们对Saccharolobus纺锤形病毒22(Ssv22)进行了基因分析，Ssv22是一种甲型杂交链病毒，其开放阅读框架(ORF)比大多数分离的杂交链病毒少。缺失和移码突变都被引入到病毒基因组的几乎所有26个开放阅读框中。总共有17个开放阅读框是必不可少的，另外还需要两个开放阅读框才能使病毒具有最佳的感染力。VP2或VP3这两种结构蛋白的缺失不会影响病毒的形态或感染性。获得了一个最小基因组长度为20个ORF的感染性Ssv22衍生物。SSV22衣壳能够容纳比野生型病毒大18kb的∼或7kb的∼。病毒衣壳的长度和宽度都随着基因组的大小而变化，但形状并不相同。我们的结果将有助于分析Ssv22在病毒感染过程中与宿主之间关键的蛋白质-蛋白质相互作用，并有助于探索Ssv22作为DNA载体在潜在应用中的用途。
A large number of fuselloviruses have been found in acidic hot springs around the globe. They share a set of highly conserved genes (core genes) and possess a varying number of less-conserved genes (non-core genes). However, the functions of most of these genes are unknown. Recent studies show that as many as half of these genes tolerate mutation. In this study, we conducted a genetic analysis on Saccharolobus spindle-shaped virus 22 (SSV22), an alphafusellovirus with fewer open reading frames (ORFs) than most of the isolated fuselloviruses. Both deletion and frame-shift mutations were introduced into nearly all of the 26 ORFs of the viral genome. A total of 17 ORFs were indispensable, and two additional ORFs were required for the optimal infectivity of the virus. Deletion of either VP2 or VP3, the two structural proteins, did not affect the morphology or infectivity of the virus. An infectious SSV22 derivative carrying a minimal genome of 20 ORFs was obtained. The SSV22 capsid was capable of accommodating a genome as large as ∼18 kb, or ∼7 kb larger than that of the wild-type virus. The viral capsid varied in both the length and width, but not in shape, with the size of the genome. Our results will facilitate the analysis of crucial protein-protein interactions between SSV22 and the host during viral infection and help explore the use of SSV22 as a vector for DNA delivery in potential applications.