| 109 | 0 | 712 |
| 下载次数 | 被引频次 | 阅读次数 |
探究非洲猪瘟病毒（ASFV）RNA解旋酶A859L的基本生物学特征，为后续研究A859L的生物学功能以及A859L在ASFV复制和致病机制中的作用奠定基础。首先，在NCBI Virus数据库中下载不同ASFV毒株的A859L基因序列，构建系统进化树。其次，利用在线软件分析A859L的理化性质和结构。最后，通过构建A859L的真核表达质粒，利用Western-blot和间接免疫荧光试验检测A859L的细胞定位和结合dsRNA的能力。系统进化树显示，A859L在不同ASFV毒株基因组中是一个保守基因，且A859L是一个亲水且不含有跨膜结构域的病毒蛋白。A859L蛋白二级结构中含有α-螺旋（49.65%）、无规则卷曲（37.88%）和延伸链（10.14%）。Western-blot和间接免疫荧光试验结果显示，A859L蛋白主要定位于细胞质且具有结合dsRNA的功能。本研究证实A859L是一个保守基因，揭示A859L蛋白定位于细胞质且具有结合dsRNA的能力，为后续探究A859L蛋白在基因组转录和复制中的作用奠定了基础。
Abstract：The aim is to investigate the basic biological characteristics of the African swine fever virus(ASFV) RNA helicase A859L,laying the foundation for subsequent studies of its biological functions and its role in ASFV replication and pathogenesis.First,the A859L gene sequences of different ASFV strains were downloaded from the NCBI Virus database,and a phylogenetic tree was constructed.Second,the physicochemical properties and structure of A859L were analyzed using online software.Finally,an eukaryotic expression plasmid for A859L was constructed,and the cellular localization and ds RNA-binding ability of A859L were detected through Western-blot and indirect immunofluorescence assay.The phylogenetic tree revealed that A859L was a conserved gene in the genomes of various ASFV strains,and A859L was a hydroph ilic viral protein without a transmembrane domain.The secondary structure of the A859L protein contains α-helices(49.65%),random coils(37.88%),and extended chains(10.14%).Western-blot and indirect immunofluorescence assay results showed that the A859L protein was primarily localized in the cytoplasm and had the ability to bind ds RNA.In conclusion:this study confirms that A859L is a conserved gene,reveals the cytoplasmic localization of the A859L protein and its ds RNA-binding capability,and provides a foundation for further exploring the role of the A859L protein in genome transcription and replication.
[1]ZHENG Y X,LI S,LI S H,et al.Transcriptome profiling in swine macrophages infected with African swine fever virus at single-cell resolution[J].Proc Natl Acad Sci USA,2022,119(19):e2201288119.
[2]URBANO A C,FERREIRA F.African swine fever control and prevention:an update on vaccine development[J].Emerg Microbes Infect,2022,11(1):2021-2033.
[3]YANG K D,XUE Y,NIU H,et al.African swine fever virus MGF360-11L negatively regulates cGAS-STINGmediated inhibition of typeⅠinterferon production[J].Vet Res,2022,53(1):7.
[4]GE S,LI J,FAN X,et al.Molecular characterization of African swine fever virus,China,2018[J].Emerg Infect Dis,2018,24(11):2131-2133.
[5]WANG N,ZHAO D M,WANG J L,et al.Architecture of African swine fever virus and implications for viral assembly[J].Science,2019,366(6465):640-644.
[6]WANG T ,LUO R ,ZHANG J ,et al.The MGF300-2R protein of African swine fever virus is associated with viral pathogenicity by promoting the autophagic degradation of IKKα and IKKβ through the recruitment of TOLLIP[J].PLoS Pathog ,2023 ,19(8):e1011580.
[7]LIU Y N,SHEN Z,XIE Z H,et al.African swine fever virus I73R is a critical virulence-related gene:A potential target for attenuation[J].Proc Natl Acad Sci USA,2023,120(15):e2210808120.
[8]RICHT J A,RAN Y,LI D,et al.African swine fever virus I267L acts as an important virulence factor by inhibiting RNA polymeraseⅢ-RIG-Ⅰ-mediated innate immunity[J].PLo S Pathog,2022,18(1):e1010-270.
[9]LI G,FU D,ZHANG G,et al.Crystal structure of the African swine fever virus structural protein p35reveals its role for core shell assembly[J].Protein Cell,2020,11(8):600-605.
[10]HONDELE M,SACHDEV R,HEINRICH S,et al.DEAD-box ATPases are global regulators of phase-separated organelles[J].Nature,2019,573(7772):144-148.
[11]JIANG C,TANG Y,DING L,et al.Targeting the N terminus of eIF4AI for inhibition of its catalytic recycling[J].Cell Chem Biol,2019,26(10):1417-1426.e5.
[12]FREITAS F B,FROUCO G,MARTINS C,et al.The QP509L and Q706L superfamilyⅡRNA helicases of African swine fever virus are required for viral replication,having non-redundant activities[J].Emerg Microbes Infect,2019,8(1):291-302.
[13]RAMIREZ-MEDINA E,VUONO E A,PRUITT S,et al.Evaluation of an ASFV RNA helicase gene A859L for virus replication and swine virulence[J].Viruses,2022,14(1):10.
[14]LIU S,LUO Y,WANG Y,et al.Cryo-EM structure of the African swine fever virus[J].Cell Host Microbe,2019,26(6):836-843.e3.
[15]VU H L X,Mc VEY D S.Recent progress on gene-deleted live-attenuated African swine fever virus vaccines[J].NPJ Vaccines,2024,9(1):60.
[16]ZHANG K S,YANG B,SHEN C C,et al.MGF360-9L is a major virulence factor associated with the African swine fever virus by antagonizing the JAK/STAT signaling pathway[J].m Bio,2022,13(1):e0233021.
[17]LIU Y,XIE Z,LI Y,et al.Evaluation of an I177L genebased five-gene-deleted African swine fever virus as a live attenuated vaccine in pigs[J].Emerg Infect Dis,2022,12(1):2148560.
[18]BAI L ,DONG J Z ,LIU Z Q ,et al.Viperin catalyzes methionine oxidation to promote protein expression and function of helicases[J].Sci Adv,2019,5(8):eaax1031.
[19]SHU T,HUANG M,WU D,et al.SARS-coronavirus-2 Nsp13 possesses NTPase and RNA helicase activities that can be inhibited by bismuth salts[J].Virol Sin,2020,35(3):321-329.
[20]ZHAO R,SHU W,HU W S,et al.Structure-based discovery of dual-target inhibitors of the helicase from Bagaza virus[J].Int J Biol Macromol,2025,294:139536.
基本信息:
DOI：10.16656/j.issn.1673-4696.2026.0024
中图分类号:S852.651
引用信息:
[1]周万卉,武正前,张旭辉,等.非洲猪瘟病毒解旋酶A859L的生物学特征[J].中国兽医科学,2026,56(02):193-199.DOI:10.16656/j.issn.1673-4696.2026.0024.
基金信息:
甘肃省高校教师创新基金项目(2025B-064); 动物疫病防控全国重点实验室开放基金项目(SKLADCPKFKT202408)
2025-09-30
2025-09-30
2025-09-30