Suppr超能文献

海洋病毒性出血性败血症病毒RNA聚合酶的单个氨基酸突变(I1012F)改变了对虹鳟鳃上皮细胞的体外毒力。

A single amino acid mutation (I1012F) of the RNA polymerase of marine viral hemorrhagic septicemia virus changes in vitro virulence to rainbow trout gill epithelial cells.

作者信息

Kim Sung-Hyun, Thu Beate J, Skall Helle F, Vendramin Niccolò, Evensen Oystein

机构信息

Norwegian University of Life Sciences, Oslo, Norway.

Department of Animal Science, Aarhus University, Aarhus, Denmark.

出版信息

J Virol. 2014 Jul;88(13):7189-98. doi: 10.1128/JVI.00423-14. Epub 2014 Apr 9.

Abstract

UNLABELLED

Viral hemorrhagic septicemia virus (VHSV) is separated into four different genotypes (I to IV) with different sublineages (K. Einer-Jensen, P. Ahrens, R. Forsberg, and N. Lorenzen, J. Gen. Virol. 85:1167-1179, 2004; K. Einer-Jensen, J. Winton, and N. Lorenzen, Vet. Microbiol. 106:167-178, 2005). European marine VHSV strains (of genotypes I to III) are, in general, nonpathogenic or have very low pathogenicity to rainbow trout after a waterborne challenge, and here we also show that genotype IVa is nonpathogenic to trout. Despite several attempts, it has not been possible to link genomic variation to in vivo virulence. In vitro virulence to gill epithelial cells (GECs) has been used as a proxy for in vivo virulence, and here we extend these studies further with the purpose of identifying residues associated with in vitro virulence. Genotype Ia (DK-3592B) and III (NO/650/07) isolates, which are pathogenic to rainbow trout (O. B. Dale, I. Orpetveit, T. M. Lyngstad, S. Kahns, H. F. Skall, N. J. Olesen, and B. H. Dannevig, Dis. Aquat. Organ. 85:93-103, 2009), were compared to two marine strains that are nonpathogenic to trout, genotypes Ib (strain 1p8 [H. F. Mortensen, O. E. Heuer, N. Lorenzen, L. Otte, and N. J. Olesen, Virus Res. 63:95-106, 1999]) and IVa (JF-09). DK-3592 and NO/650/07 were pathogenic to GECs, while marine strains 1p8 and JF-09 were nonpathogenic to GECs. Eight conserved amino acid substitutions contrasting high- and low-virulence strains were identified, and reverse genetics was used in a gain-of-virulence approach based on the JF-09 backbone. Mutations were introduced into the G, NV, and L genes, and seven different virus clones were obtained. For the first time, we show that a single amino acid mutation in conserved region IV of the L protein, I1012F, rendered the virus able to replicate and induce a cytopathic effect in trout GECs. The other six mutated variants remained nonpathogenic.

IMPORTANCE

This is the first study to clearly link in vitro virulence of viral hemorrhagic septicemia virus (VHSV) with an amino acid residue in the L protein, a site located in conserved region IV of the L protein. In vitro virulence is documented by induction of cytopathic effects and viability studies of gill epithelial cells, and the observed cellular responses to infection are associated with increased viral replication levels. There are no previous studies addressing the importance of the L protein or the RNA-dependent RNA polymerase for virus virulence in vitro or in vivo. Therefore, the findings reported here should broaden the search for pathogenicity traits in novirhabdoviruses, and there is a possibility that the polymerase participates in defining the host species virulence of various VHSV strains.

摘要

未标记

病毒性出血性败血症病毒(VHSV)分为四种不同的基因型(I至IV),具有不同的亚系(K. Einer-Jensen、P. Ahrens、R. Forsberg和N. Lorenzen,《普通病毒学杂志》85:1167 - 1179,2004年;K. Einer-Jensen、J. Winton和N. Lorenzen,《兽医微生物学》106:167 - 178,2005年)。欧洲海洋VHSV毒株(基因型I至III)一般对虹鳟鱼经水传播感染后无致病性或致病性极低,并且我们在此还表明IVa基因型对鳟鱼无致病性。尽管进行了多次尝试,但仍无法将基因组变异与体内毒力联系起来。对鳃上皮细胞(GECs)的体外毒力已被用作体内毒力的替代指标,并且我们在此进一步扩展这些研究,目的是鉴定与体外毒力相关的残基。将对虹鳟鱼有致病性的基因型Ia(DK - 3592B)和III(NO/650/07)分离株(O. B. Dale、I. Orpetveit、T. M. Lyngstad、S. Kahns、H. F. Skall、N. J. Olesen和B. H. Dannevig,《水生生物疾病》85:93 - 103,2009年)与两种对鳟鱼无致病性的海洋毒株进行比较,即基因型Ib(毒株1p8 [H. F. Mortensen、O. E. Heuer、N. Lorenzen、L. Otte和N. J. Olesen,《病毒研究》63:95 - 106,1999年])和IVa(JF - 09)。DK - 3592和NO/650/07对GECs有致病性,而海洋毒株1p8和JF - 09对GECs无致病性。鉴定出了八个区分高毒力和低毒力毒株的保守氨基酸替换,并基于JF - 09骨架在毒力增强方法中使用了反向遗传学。将突变引入G、NV和L基因,获得了七个不同的病毒克隆。我们首次表明,L蛋白保守区域IV中的单个氨基酸突变I1012F使病毒能够在鳟鱼GECs中复制并诱导细胞病变效应。其他六个突变变体仍然无致病性。

重要性

这是第一项明确将病毒性出血性败血症病毒(VHSV)的体外毒力与L蛋白中的一个氨基酸残基联系起来的研究,该位点位于L蛋白的保守区域IV。通过诱导细胞病变效应和对鳃上皮细胞的活力研究记录了体外毒力,并且观察到的细胞对感染的反应与病毒复制水平的增加相关。以前没有研究探讨L蛋白或RNA依赖性RNA聚合酶在体外或体内对病毒毒力的重要性。因此,此处报道的发现应拓宽对新弹状病毒致病性特征的研究范围,并且聚合酶有可能参与定义各种VHSV毒株的宿主物种毒力。

相似文献

引用本文的文献

1
Transcriptome Profiling of Infected with Low or Highly Pathogenic Viral Hemorrhagic Septicemia Virus (VHSV).
Microorganisms. 2023 Dec 28;12(1):57. doi: 10.3390/microorganisms12010057.
3
Naturally occurring substitution in one amino acid in VHSV phosphoprotein enhances viral virulence in flounder.
PLoS Pathog. 2021 Jan 19;17(1):e1009213. doi: 10.1371/journal.ppat.1009213. eCollection 2021 Jan.
6
The (VHSV) Markers of Virulence in Rainbow Trout ().
Front Microbiol. 2020 Oct 20;11:574231. doi: 10.3389/fmicb.2020.574231. eCollection 2020.
8
VHSV Single Amino Acid Polymorphisms (SAPs) Associated With Virulence in Rainbow Trout.
Front Microbiol. 2020 Aug 27;11:1984. doi: 10.3389/fmicb.2020.01984. eCollection 2020.

本文引用的文献

5
Nonvirion protein of novirhabdovirus suppresses apoptosis at the early stage of virus infection.
J Virol. 2011 Aug;85(16):8393-402. doi: 10.1128/JVI.00597-11. Epub 2011 Jun 8.
8
Molecular architecture of the vesicular stomatitis virus RNA polymerase.
Proc Natl Acad Sci U S A. 2010 Nov 16;107(46):20075-80. doi: 10.1073/pnas.1013559107. Epub 2010 Nov 1.
9
A reverse genetics system for the Great Lakes strain of viral hemorrhagic septicemia virus: the NV gene is required for pathogenicity.
Mar Biotechnol (NY). 2011 Aug;13(4):672-83. doi: 10.1007/s10126-010-9329-4. Epub 2010 Oct 9.
10
Alpha interferon and not gamma interferon inhibits salmonid alphavirus subtype 3 replication in vitro.
J Virol. 2010 Sep;84(17):8903-12. doi: 10.1128/JVI.00851-10. Epub 2010 Jun 23.

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验