Characterization of an envelope protein 118L in invertebrate iridescent virus 6 (IIV6).

IF 1.9 4区 医学 Q3 GENETICS & HEREDITY
Virus Genes Pub Date : 2024-10-01 Epub Date: 2024-06-26 DOI:10.1007/s11262-024-02082-7
Betul Altun, Kubra Zengin, Sevde Yayli Dabag, Aydin Yesilyurt, Remziye Nalcacioglu, Zihni Demirbag
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Abstract

Invertebrate iridescent virus 6 (IIV6) is a nucleocytoplasmic insect virus and a member of the family Iridoviridae. The IIV6 genome consists of 212,482 bp of linear dsDNA with 215 non-overlapping and putative protein-encoding ORFs. The IIV6 118L ORF is conserved in all sequenced members of the Iridoviridae and encodes a 515 amino acid protein with three predicted transmembrane domains and several N-glycosylation/N-myristoylation sites. In this study, we characterized the 118L ORF by both deleting it from the viral genome and silencing its expression with dsRNA in infected insect cells. The homologous recombination method was used to replace 118L ORF with the green fluorescent protein (gfp) gene. Virus mutants in which the 118L gene sequence had been replaced with gfp were identified by fluorescence microscopy but could not be propagated separately from the wild-type virus in insect cells. Unsuccessful attempts to isolate the mutant virus with the 118L gene deletion suggested that the protein is essential for virus replication. To support this result, we used dsRNA to target the 118L gene and showed that treatment resulted in a 99% reduction in virus titer. Subsequently, we demonstrated that 118L-specific antibodies produced against the 118L protein expressed in the baculovirus vector system were able to neutralize the virus infection. All these results indicate that 118L is a viral envelope protein that is required for the initiation of virus replication.

Abstract Image

无脊椎动物虹彩病毒 6(IIV6)包膜蛋白 118L 的特征。
无脊椎动物虹彩病毒 6(IIV6)是一种核细胞质昆虫病毒,属于虹彩病毒科。IIV6 基因组由 212,482 bp 的线性 dsDNA 组成,其中有 215 个不重叠的假定蛋白编码 ORF。IIV6 118L ORF 在所有已测序的ridoviridae成员中都是保守的,它编码一种 515 氨基酸的蛋白质,具有三个预测的跨膜结构域和几个 N-糖基化/N-肉豆蔻酰化位点。在本研究中,我们通过从病毒基因组中删除 118L ORF 和用 dsRNA 在受感染的昆虫细胞中沉默其表达来鉴定 118L ORF 的特征。用同源重组法将 118L ORF 替换为绿色荧光蛋白(gfp)基因。用荧光显微镜鉴定了用 gfp 基因替换了 118L 基因序列的病毒突变体,但无法在昆虫细胞中与野生型病毒分开繁殖。分离 118L 基因缺失突变体病毒的尝试未获成功,这表明该蛋白对病毒复制至关重要。为了支持这一结果,我们使用 dsRNA 靶向 118L 基因,结果表明处理后病毒滴度降低了 99%。随后,我们证明了针对在杆状病毒载体系统中表达的 118L 蛋白产生的 118L 特异性抗体能够中和病毒感染。所有这些结果表明,118L 是一种病毒包膜蛋白,是病毒复制启动所必需的。
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来源期刊
Virus Genes
Virus Genes 医学-病毒学
CiteScore
3.30
自引率
0.00%
发文量
76
审稿时长
3 months
期刊介绍: Viruses are convenient models for the elucidation of life processes. The study of viruses is again on the cutting edge of biological sciences: systems biology, genomics, proteomics, metagenomics, using the newest most powerful tools. Huge amounts of new details on virus interactions with the cell, other pathogens and the hosts – animal (including human), insect, fungal, plant, bacterial, and archaeal - and their role in infection and disease are forthcoming in perplexing details requiring analysis and comments. Virus Genes is dedicated to the publication of studies on the structure and function of viruses and their genes, the molecular and systems interactions with the host and all applications derived thereof, providing a forum for the analysis of data and discussion of its implications, and the development of new hypotheses.
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