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The role of viral protein Ac34 in nuclear relocation of subunits of the actin-related protein 2/3 complex

Virologica Sinica 2016年第6期31卷 480-489页

作者： jingfang mu Yongli Zhang Yangyang Hu Xue Hu Yuan Zhou Xinwen Chen Yun Wang State Key Laboratory of Virology Wuhan Institute of Virology Chinese Academy of Sciences Wuhan 430071 China University of Chinese Academy of Sciences Beijing 100049 China

The actin nucleator actin-related protein complex(Arp2/3) is composed of seven subunits: Arp2,Arp3, p40/ARPC1(P40), p34/ARPC2(P34), p21/ARPC3(P21), p20/ARPC4(P20), and p16/ARPC5(P16). Arp2/3 plays crucial roles in a variety of cellular activities through regulation of actin polymerization. Autographa californica multiple nucleopolyhedrovirus(Ac MNPV), one of the beststudied alphabaculoviruses, induces Arp2/3 nuclear relocation and mediates nuclear actin polymerization to assist in virus replication. We have demonstrated that Ac34, a viral late-gene product, induces translocation of the P40 subunit of Arp2/3 to the nucleus during Ac MNPV infection. However, it remains unknown whether Ac34 could relocate other Arp2/3 subunits to the nucleus. In this study, the effects of the viral protein Ac34 on the distribution of these subunits were studied by an immunofluorescence assay. Arp2, P34, P21, and P20 cloned from Spodoptera frugiperda(Sf9) cells showed mainly cytoplasmic localization and were relocated to the nucleus in the presence of Ac34. In addition, Arp3 was localized in the cytoplasm in both the presence and absence of Ac34, and P16 showed whole-cell localization. In contrast to Sf9 cells, all subunits of mammalian Arp2/3 showed no nuclear relocation in the presence of Ac34. Co-immunoprecipitation analysis of the interaction between Ac34 and Arp2/3 subunits revealed that Ac34 bound to P40,P34, and P20 of Sf9 cells. However, none of the subunits of mammalian Arp2/3 interacted with Ac34, indicating that protein-protein interaction is essential for Ac34 to relocate Arp2/3 subunits to the nucleus.

关键词： Autographa californica multiple nucleopolyhedrovirus(AcMNPV) Ac34 Arp2/3 complex nuclear relocation

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Hepatitis C Virus NS2 Protein Suppresses RNA Interference in Cells

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Virologica Sinica 2020年第4期35卷 436-444页

作者： Hui Zhou Qi Qian Ting Shu Jiuyue Xu Jing Kong jingfang mu Yang Qiu Xi Zhou State Key Laboratory of Virology College of Life SciencesWuhan UniversityWuhan 430072China State Key Laboratory of Virology Wuhan Institute of VirologyCenter for Biosafety Mega-ScienceChinese Academy of SciencesWuhan 430071China Center for Translational Medicine Wuhan Jinyintan HospitalWuhan 430040China University of Chinese Academy of Sciences Beijing 100049China

RNAi interference(RNAi)is an evolutionarily conserved post-transcriptional gene silencing mechanism and has been well recognized as an important antiviral immunity in *** viruses have been shown to encode viral suppressors of RNAi(VSRs)to antagonize antiviral *** C virus(HCV)is a medically important human pathogen that causes acute and chronic *** this study,we screened all the nonstructural proteins of HCV and found that HCV NS2 could suppress RNAi induced either by small hairpin RNAs(shRNAs)or small interfering RNAs(siRNAs)in mammalian ***,we demonstrated that NS2 could suppress RNAi via its direct interaction with doublestranded RNAs(dsRNAs)and siRNAs,and further identified that the cysteine 184 of NS2 is required for the RNAi suppression activity through a serial of point mutation ***,our findings uncovered that HCV NS2 can act as a VSR in vitro,thereby providing novel insights into the life cycle and virus-host interactions of HCV.

关键词： Hepatitis C virus(HCV) NS2 Antiviral RNAi Viral suppressor of RNAi(VSR)

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Mechanisms and Effects on HBV Replication of the Interaction between HBV Core Protein and Cellular Filamin B

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Virologica Sinica 2018年第2期33卷 162-172页

作者： Yilin Li Yishuang Sun Fuyun Sun Rong Hua Chenlin Li Lang Chen Deyin Guo jingfang mu School of Basic Medical Sciences Wuhan University Wuhan 430071 China State Key Laboratory of Virology Wuhan Institute of Virology Chinese Academy of Sciences Wuhan 430071 China School of Basic Medicine (Shenzhen) Sun Yat-sen University Guangzhou 510081 China

Hepatitis B virus(HBV) infection is one of the major problems that threatens global health. There have been many studies on HBV, but the relationship between HBV and host factors is largely unexplored and more studies are needed to clarify these interactions. Filamin B is an actin-binding protein that acts as a cytoskeleton protein, and it is involved in cell development and several signaling pathways. In this study, we showed that filamin B interacted with HBV core protein,and the interaction promoted HBV replication. The interaction between filamin B and core protein was observed in HEK293T, Huh7 and HepG2 cell lines by co-immunoprecipitation and co-localization immnofluoresence. Overexpression of filamin B increased the levels of HBV total RNAs and pre-genome RNA(pg RNA), and improved the secretion level of hepatitis B surface antigen(HBsAg) and hepatitis B e antigen(HBeAg). In contrast, filamin B knockdown inhibited HBV replication, decreased the level of HBV total RNAs and pgRNA, and reduced the secretion level of HBsAg and HBeAg. In addition, we found that filamin B and core protein may interact with each other via four blocks of argentine residues at the C-terminus of core protein. In conclusion, we identify filamin B as a novel host factor that can interact with core protein to promote HBV replication in hepatocytes. Our study provides new insights into the relationship between HBV and host factors and may provide new strategies for the treatment of HBV infection.

关键词： Filamin B Core HBV replication

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SARS-Coronavirus-2 Nsp13 Possesses NTPase and RNA Helicase Activities That Can Be Inhibited by Bismuth Salts

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Virologica Sinica 2020年第3期35卷 321-329页

作者： Ting Shu muhan Huang Di Wu Yujie Ren Xueyi Zhang Yang Han jingfang mu Ruibing Wang Yang Qiu Ding-Yu Zhang Xi Zhou Center for Translational Medicine Wuhan Jinyintan HospitalWuhan 430023China State Key Laboratory of Virology Wuhan Institute of VirologyCenter for Biosafety Mega-ScienceChinese Academy of SciencesWuhan 430071China Center for Precision Translational Medicine of Wuhan Institute of Virology and Guangzhou Women and Children’s Medical Center Guangzhou Women and Children’s Medical CenterGuangzhou 510120China State Key Laboratory of Quality Research in Chinese Medicine Institute of Chinese Medical SciencesUniversity of MacaoMacao SAR 999078China University of Chinese Academy of Sciences Beijing 100049China

The ongoing outbreak of Coronavirus Disease 2019(COVID-19)has become a global public health ***-2(SARS-CoV-2),the causative pathogen of COVID-19,is a positive-sense single-stranded RNA virus belonging to the family *** 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 ***,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 ***,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 ***,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.

关键词： SARS-coronavirus-2(SARS-CoV-2) Nsp13 NTPase Helicase Antiviral target

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The ORF3a protein of SARS-CoV-2 induces apoptosis in cells

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Cellular & Molecular Immunology 2020年第8期17卷 881-883页

作者： Yujie Ren Ting Shu Di Wu jingfang mu Chong Wang muhan Huang Yang Han Xue-Yi Zhang Wei Zhou Yang Qiu Xi Zhou Center for Precision Translational Medicine of Wuhan Institute of Virology&Guangzhou Women and Children’s Medical Center Guangzhou Women and Children’s Medical CenterGuangzhouGuangdong510120China State Key Laboratory of Virology Wuhan Institute of VirologyCenter for Biosafety Mega-ScienceChinese Academy of Sciences(CAS)WuhanHubei430071China Center for Precision Translational Medicine of Wuhan Institute of Virology&Guangzhou Women and Children’s Medical Center Wuhan Institute of VirologyCASWuhanHubei430071China Joint Laboratory of Infectious Diseases and Health Wuhan Institute of Virology&Wuhan Jinyintan HospitalCASWuhanHubei430023China Center for Translational Medicine Wuhan Jinyintan HospitalWuhanHubei430023China State Key Laboratory of Virology College of Life SciencesWuhan UniversityWuhanHubei430072China University of Chinese Academy of Sciences Beijing100049China

Severe acute respiratory syndrome coronavirus-2(SARS-CoV-2)has caused the ongoing pandemic of Coronavirus Disease ***-CoV-2 belongs to the genus Betacoronavirus of the Coronaviridae family,which includes SARS-CoV and Middle East respiratory syndrome coronavirus.1,2 Coronavirus-encoded accessory proteins play critical roles in virus–host interactions and the modulation of host immune responses,thereby contributing to coronaviral pathogenicity via different strategies.3 However,the functions of SARS-CoV-2-encoded accessory proteins are not well *** is a predominant type of programmed cell death,and has been recognized as an important host antiviral defense mechanism that controls viral infection and regulates the inflammatory response.4,5 Previous studies have reported that the SARS-CoV-encoded accessory protein ORF3a can induce apoptosis in cells,6,7 leading to the question of whether SARS-CoV-2 ORF3a also has pro-apoptotic ***,we investigated the potential apoptosis-inducing activity of SARS-CoV-2 ORF3a in different cell lines and compared the pro-apoptotic activities of SARS-CoV-2 ORF3a with those of SARS-CoV ORF3a using the same system.

关键词： respiratory acute thereby

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SARS-CoV-2-encoded nucleocapsid protein acts as a viral suppressor of RNA interference in cells

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Science China(Life Sciences) 2020年第9期63卷 1413-1416页

作者： jingfang mu Jiuyue Xu Leike Zhang Ting Shu Di Wu muhan Huang Yujie Ren Xufang Li Qing Geng Yi Xu Yang Qiu Xi Zhou State Key Laboratory of Virology Wuhan Institute of VirologyCenter for Biosafety Mega-ScienceChinese Academy of Sciences(CAS)Wuhan 430071China Joint Laboratory of Infectious Diseases and Health Wuhan Institute of Virology&Wuhan Jinyintan HospitalCASWuhan 430023China The University of Chinese Academy of Sciences Beijing 100049China Center for Translational Medicine Wuhan Jinyintan HospitalWuhan 430040China Center for Precision Translational Medicine of Wuhan Institute of Virology and Guangzhou Women and Children’s Medical Center Guangzhou Women and Children’s Medical CenterGuangzhou 510120China Department of Thoracic Surgery Renmin Hospital of Wuhan UniversityWuhan 430060China

Dear Editor,Coronaviruses (Co Vs) are large enveloped non-segmented positive-strand RNA viruses that broadly distribute among humans and other animal species, including bats, mice and birds. SARS-Co V-2 infections can cause diseases, named the 2019 novel coronavirus disease (COVID-19). The symptoms of COVID-19 range from mild symptoms to severe respiratory syndromes, including pneumonia, and even death(Chen et al., 2020b;Jiang and Shi, 2020;Xia et al., 2020).

关键词： al. suppressor diseases

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Reviving chloroquine for anti-SARS-CoV-2 treatment with cucurbit[7]uril-based supramolecular formulation

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Chinese Chemical Letters 2021年第10期32卷 3019-3022页

作者： Cheryl H.T.Kwong jingfang mu Shengke Li Yaohui Fang Qianyun Liu Xiangjun Zhang Hiotong Kam Simon M.Y.Lee Yu Chen Fei Deng Xi Zhou Ruibing Wang State Key Laboratory of Quality Research in Chinese Medicine Institute of Chinese Medical SciencesUniversity of MacaoMacao 999078China State Key Laboratory of Virology Wuhan Institute of VirologyCenter for Biosafety Mega-ScienceChinese Academy of Sciences(CAS)Wuhan 430071China State Key Laboratory of Virology College of Life SciencesWuhan UniversityWuhan 430072China

The wide-spreading SARS-CoV-2 virus has put the world into boiling water for more than a year,however pharmacological therapies to act effectively against coronavirus disease 2019(COVID-19)remain ***(CQ),an antimalarial drug,was found to exhibit promising antiviral activity in vitro and in vivo at a high dosage,thus CQ was approved by the FDA for the emergency use authorization(EUA)in the fight against COVID-19 in the US,but later was revoked the EUA status due to the severe clinical ***,we show that supramolecular formulation of CQ by a macrocyclic host,curcurbit[7]uril(CB[7]),reduced its non-specific toxicity and improved its antiviral activity against coronavirus,working in synergy with CB[7].CB[7]was found to form 1:1 host-guest complexes with CQ,with a binding constant of$104 L/*** CQ-CB[7]formulation decreased the cytotoxicity of CQ against Vero E6 and L-02 cell *** particular,the cytotoxicity of CQ(60 mmol/L)against both Vero E6 cell line and L-02 cell lines was completely inhibited in the presence of 300 mmol/L and 600 mmol/L CB[7],***,the CB[7]alone showed astonishing antiviral activity in SARS-CoV-2 infected Vero E6 cells and mouse hepatitis virus strain A59(MHV-A59)infected N2 A cells,and synergistically improved the antiviral activity of CQ-CB[7],suggesting that CB[7]-based CQ formulation has a great potential as a safe and effective antiviral agent against SARS-CoV-2 and other coronavirus.

关键词： Host-guest Chloroquine Cucurbituril SARS-CoV-2 COVID-19

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Plasma metabolomic and lipidomic alterations associated with COVID-19

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National Science Review 2020年第7期7卷 1157-1168页

作者： Di Wu Ting Shu Xiaobo Yang Jian-Xin Song Mingliang Zhang Chengye Yao Wen Liu muhan Huang Yuan Yu Qingyu Yang Tingju Zhu Jiqian Xu jingfang mu Yaxin Wang Hong Wang Tang Tang Yujie Ren Yongran Wu Shu-Hai Lin Yang Qiu Ding-Yu Zhang You Shang Xi Zhou Joint Laboratory of Infectious Diseases and Health Wuhan Institute of Virology & Wuhan Jinyintan Hospital Wuhan Institute of Virology Center for Biosafety Mega-ScienceChinese Academy of Sciences (CAS) State Key Laboratory of Virology Wuhan Institute of VirologyCenter for Biosafety Mega-Science CAS Center for Translational Medicine Jinyintan Hospital Joint Laboratory of Infectious Diseases and Health Wuhan Institute of Virology & Wuhan Jinyintan Hospital Wuhan Jinyintan Hospital Department of Critical Care Medicine Union Hospital Tongji Medical College Huazhong University of Science and Technology Department of Infectious Diseases Tongji Hospital Tongji Medical College Huazhong University of Science and Technology

The pandemic of the coronavirus disease 2019(COVID-19) has become a global public health crisis. The symptoms of COVID-19 range from mild to severe, but the physiological changes associated with COVID-19 are barely understood. In this study, we performed targeted metabolomic and lipidomic analyses of plasma from a cohort of patients with COVID-19 who had experienced different symptoms. We found that metabolite and lipid alterations exhibit apparent correlation with the course of disease in these patients, indicating that the development of COVID-19 affected their whole-body metabolism. In particular, malic acid of the TCA cycle and carbamoyl phosphate of the urea cycle result in altered energy metabolism and hepatic dysfunction, respectively. It should be noted that carbamoyl phosphate is profoundly down-regulated in patients who died compared with patients with mild symptoms. And, more importantly, guanosine monophosphate(GMP), which is mediated not only by GMP synthase but also by CD39 and CD73, is significantly changed between healthy subjects and patients with COVID-19, as well as between the mild and fatal cases. In addition, dyslipidemia was observed in patients with ***, the disturbed metabolic patterns have been found to align with the progress and severity of COVID-19. This work provides valuable knowledge about plasma biomarkers associated with COVID-19 and potential therapeutic targets, as well as an important resource for further studies of the pathogenesis of COVID-19.

关键词： SARS-Co V-2 COVID-19 metabolome lipidome

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KNK437抑制乙型肝炎病毒复制及转录

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病毒学报 2017年第1期33卷 24-35页

作者：胡康洪黄亚运穆敬芳程智逵朱祥湖北工业大学生物医药研究院中德生物医学中心武汉430068 中国科学院武汉病毒研究所病毒学国家重点实验室武汉430071

乙型肝炎病毒(Hepatitis B virus,HBV)在肝细胞内复制过程中,病毒反转录酶(P蛋白)与前基因组RNA(Pregenomic RNA,pgRNA)上的RNA包装信号ε形成P-ε复合物,启动反转录合成和核衣壳装配。封闭P-ε相互作用是一种极具吸引力的抗HBV策略。已... 详细信息

乙型肝炎病毒(Hepatitis B virus,HBV)在肝细胞内复制过程中,病毒反转录酶(P蛋白)与前基因组RNA(Pregenomic RNA,pgRNA)上的RNA包装信号ε形成P-ε复合物,启动反转录合成和核衣壳装配。封闭P-ε相互作用是一种极具吸引力的抗HBV策略。已知P-ε形成RNP复合物正常发挥活性时,需要热休克蛋白(Heat-shock proteins,Hsps)参与。本研究探索Hsps抑制剂KNK437对HBV复制及转录的影响。主要运用了三种工作模型:HepG2.2.15细胞系、瞬时转染1.05×HBV(pCH9-3091)质粒的Huh7细胞系和瞬时转染1.3×HBV(pGEM-1.3×HBV)质粒的Huh7细胞系。采用CCK-8方法检测KNK437的细胞毒性,ELISA测定细胞培养液上清中HBsAg和HBeAg的分泌水平;q-PCR和qRT-PCR分别检测细胞内HBV核衣壳中DNA和细胞内HBV RNA水平;Western blotting检测细胞内衣壳亚单位(core)表达水平;qRT-PCR检测细胞内Hsps本身的转录水平。结果表明:20μM KNK437对细胞没有毒性,且在该浓度下KNK437除HepG2.2.15模型外,均下调HBV HBsAg和HBeAg的胞外分泌,抑制细胞内HBV核衣壳中DNA合成,降低细胞内HBV RNA转录水平,最低可使DNA水平降至1.5%左右,RNA水平降至30%左右,从而表明KNK437抑制HBV复制和转录。在HepG2.2.15中的Western blotting显示,KNK437明显减少细胞内衣壳亚单位(core)表达水平。KNK437也抑制Hsp70,Hsp90b,Hsp40等三种热休克蛋白RNA的转录,从而验证了它的确是一种泛Hsps抑制剂。本研究结果显示KNK437具有潜在抗HBV应用前景。

关键词：乙型肝炎病毒(HBV) 热休克蛋白(Hsps) KNK437 抑制剂

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生活垃圾焚烧厂烟气中二■英的分布及净化特征

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环境化学 2019年第10期38卷 2256-2265页

作者：杨虎城陈柏杭牟靖芳叶红艺李世平徐欢刘敬勇广州环保投资集团有限公司广州510030 广州普诺环境检测技术有限公司广州510700 广东工业大学环境科学与工程学院广州510006

以广州市某大型生活垃圾焚烧发电厂的3台焚烧炉(750 t·d^-1)为对象,研究了焚烧烟气净化前后二噁英类同系物的分布特征及其污染控制效果.结果表明,采用“半干法脱酸+活性炭+布袋除尘”烟气净化工艺对二噁英的去除率能达到97.3%—99.4%.... 详细信息

以广州市某大型生活垃圾焚烧发电厂的3台焚烧炉(750 t·d^-1)为对象,研究了焚烧烟气净化前后二噁英类同系物的分布特征及其污染控制效果.结果表明,采用“半干法脱酸+活性炭+布袋除尘”烟气净化工艺对二噁英的去除率能达到97.3%—99.4%.焚烧烟气中二噁英类同系物的分布特征表明,生活垃圾焚烧发电过程中二噁英主要在烟气换热过程合成.烟气净化过程中,活性炭喷射+布袋除尘净化系统对二噁英类同系物的去除效率存在一个优先级关系,即PCDDs〉PCDFs且高氯代PCDFs〉低氯代PCDFs.净化后的烟气中二噁英类同系物2,3,4,7,8-PeCDF的质量浓度与对应的二噁英当量浓度的相关性最高,相关系数R 2为0.959,可以考虑将2,3,4,7,8-PeCDF作为二噁英的快速应急检测的指示物.

关键词：生活垃圾焚烧二噁英类同系物烟气排放

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