The ongoing outbreak of Coronavirus Disease 2019(COVID-19)has become a global public health emergency.SARScoronavirus-2(SARS-CoV-2),the causative pathogen of COVID-19,is a positive-sense single-stranded RNA virus belo...The ongoing outbreak of Coronavirus Disease 2019(COVID-19)has become a global public health emergency.SARScoronavirus-2(SARS-CoV-2),the causative pathogen of COVID-19,is a positive-sense single-stranded RNA virus belonging to the family Coronaviridae.For RNA viruses,virus-encoded RNA helicases have long been recognized to play pivotal roles during viral life cycles by facilitating the correct folding and replication of viral RNAs.Here,our studies show that SARS-CoV-2-encoded nonstructural protein 13(nsp13)possesses the nucleoside triphosphate hydrolase(NTPase)and RNA helicase activities that can hydrolyze all types of NTPs and unwind RNA helices dependently of the presence of NTP,and further characterize the biochemical characteristics of these two enzymatic activities associated with SARS-CoV-2 nsp13.Moreover,we found that some bismuth salts could effectively inhibit both the NTPase and RNA helicase activities of SARS-CoV-2 nsp13 in a dose-dependent manner.Thus,our findings demonstrate the NTPase and helicase activities of SARS-CoV-2 nsp13,which may play an important role in SARS-CoV-2 replication and serve as a target for antivirals.展开更多
Background:Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)emerged in December 2019 and has led to a global coronavirus disease 2019(COVID-19)pandemic.Currently,incomplete understanding of how SARS-CoV-2 ar...Background:Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)emerged in December 2019 and has led to a global coronavirus disease 2019(COVID-19)pandemic.Currently,incomplete understanding of how SARS-CoV-2 arrogates the host cell to establish its life cycle has led to slow progress in the development of effective drugs.Results:In this study,we found that SARS-CoV-2 hijacks the host protein EWSR1(Ewing Sarcoma breakpoint region 1/EWS RNA binding protein 1)to promote the activity of its helicase NSP13 to facilitate viral propagation.NSP13 is highly conserved among coronaviruses and is crucial for virus replication,providing chemical energy to unwind viral RNA replication intermediates.Treatment with different SARS-CoV-2 NSP13 inhibitors in multi-ple cell lines infected with SARS-CoV-2 effectively suppressed SARS-CoV-2 infection.Using affinity-purification mass spectrometry,the RNA binding protein EWSR1 was then identified as a potent host factor that physically associated with NSP13.Furthermore,silencing EWSR1 dramatically reduced virus replication at both viral RNA and protein levels.Mechanistically,EWSR1 was found to bind to the NTPase domain of NSP13 and potentially enhance its dsRNA unwinding ability.Conclusions:Our results pinpoint EWSR1 as a novel host factor for NSP13 that could potentially be used for drug repurposing as a therapeutic target for COVID-19.展开更多
冠状病毒能够引发多种传染性疾病,给动物和人类的健康带来严重危害。研发有效的疫苗和抗病毒药物成为防治疾病的重要手段。冠状病毒基因组能够编码多种蛋白质,包括结构蛋白、非结构蛋白和辅助蛋白。解旋酶非结构蛋白13(nonstructural pr...冠状病毒能够引发多种传染性疾病,给动物和人类的健康带来严重危害。研发有效的疫苗和抗病毒药物成为防治疾病的重要手段。冠状病毒基因组能够编码多种蛋白质,包括结构蛋白、非结构蛋白和辅助蛋白。解旋酶非结构蛋白13(nonstructural protein 13, NSP13)是冠状病毒编码的一种关键非结构蛋白,能够调控病毒复制和宿主先天免疫反应。因此,NSP13被认为是研发抗冠状病毒药物的重要靶点。本文结合国内外现有NSP13相关研究成果,对冠状病毒解旋酶NSP13的来源与结构、序列保守性、解旋机制、酶抑制剂、蛋白互作以及免疫调控等方面进行综述,并且分析了NSP13研究目前面临的问题,为研发靶向NSP13的广谱抗冠状病毒药物提供了理论依据。展开更多
基金supported by the Strategic Priority Research Program of CAS(XDB29010300 to X.Z.)National Natural Science Foundation of China(81873964 to Y.Q.,31800140 to J.M.and 31670161 to X.Z.)+1 种基金National Science and Technology Major Project(2018ZX10101004 to X.Z.)the Science and Technology Development Fund,Macao SAR(0007/2020/A to R.W)。
文摘The ongoing outbreak of Coronavirus Disease 2019(COVID-19)has become a global public health emergency.SARScoronavirus-2(SARS-CoV-2),the causative pathogen of COVID-19,is a positive-sense single-stranded RNA virus belonging to the family Coronaviridae.For RNA viruses,virus-encoded RNA helicases have long been recognized to play pivotal roles during viral life cycles by facilitating the correct folding and replication of viral RNAs.Here,our studies show that SARS-CoV-2-encoded nonstructural protein 13(nsp13)possesses the nucleoside triphosphate hydrolase(NTPase)and RNA helicase activities that can hydrolyze all types of NTPs and unwind RNA helices dependently of the presence of NTP,and further characterize the biochemical characteristics of these two enzymatic activities associated with SARS-CoV-2 nsp13.Moreover,we found that some bismuth salts could effectively inhibit both the NTPase and RNA helicase activities of SARS-CoV-2 nsp13 in a dose-dependent manner.Thus,our findings demonstrate the NTPase and helicase activities of SARS-CoV-2 nsp13,which may play an important role in SARS-CoV-2 replication and serve as a target for antivirals.
基金This work was supported by grants from the Na-tional Science Fund for Distinguished Young Schol-ars(82025022)the Central Charity Fund of Chinese Academy of Medical Science(2020-PT310-009)+1 种基金the Sci-ence and Technology Innovation Committee of Shenzhen Municipality(2020A1111350032)the China Post-doctoral Science Foundation(2021M693359).
文摘Background:Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)emerged in December 2019 and has led to a global coronavirus disease 2019(COVID-19)pandemic.Currently,incomplete understanding of how SARS-CoV-2 arrogates the host cell to establish its life cycle has led to slow progress in the development of effective drugs.Results:In this study,we found that SARS-CoV-2 hijacks the host protein EWSR1(Ewing Sarcoma breakpoint region 1/EWS RNA binding protein 1)to promote the activity of its helicase NSP13 to facilitate viral propagation.NSP13 is highly conserved among coronaviruses and is crucial for virus replication,providing chemical energy to unwind viral RNA replication intermediates.Treatment with different SARS-CoV-2 NSP13 inhibitors in multi-ple cell lines infected with SARS-CoV-2 effectively suppressed SARS-CoV-2 infection.Using affinity-purification mass spectrometry,the RNA binding protein EWSR1 was then identified as a potent host factor that physically associated with NSP13.Furthermore,silencing EWSR1 dramatically reduced virus replication at both viral RNA and protein levels.Mechanistically,EWSR1 was found to bind to the NTPase domain of NSP13 and potentially enhance its dsRNA unwinding ability.Conclusions:Our results pinpoint EWSR1 as a novel host factor for NSP13 that could potentially be used for drug repurposing as a therapeutic target for COVID-19.
文摘冠状病毒能够引发多种传染性疾病,给动物和人类的健康带来严重危害。研发有效的疫苗和抗病毒药物成为防治疾病的重要手段。冠状病毒基因组能够编码多种蛋白质,包括结构蛋白、非结构蛋白和辅助蛋白。解旋酶非结构蛋白13(nonstructural protein 13, NSP13)是冠状病毒编码的一种关键非结构蛋白,能够调控病毒复制和宿主先天免疫反应。因此,NSP13被认为是研发抗冠状病毒药物的重要靶点。本文结合国内外现有NSP13相关研究成果,对冠状病毒解旋酶NSP13的来源与结构、序列保守性、解旋机制、酶抑制剂、蛋白互作以及免疫调控等方面进行综述,并且分析了NSP13研究目前面临的问题,为研发靶向NSP13的广谱抗冠状病毒药物提供了理论依据。
