A型流感病毒RNA聚合酶及其对基因组复制和转录的调控作用

A型流感病毒是正粘病毒科成员,为单股负链分节段RNA病毒,全基因组由八个节段组成,分别编码八种结构蛋白(PB2、PB1、PA、HA、NP、NA、M1和M2)和两种非结构蛋白(NS1和NS2)。核蛋白(NP)和RNA聚合酶复合体与病毒的八个RNA节段组成八个螺旋丝状的病毒核衣壳(RNP),核衣壳被双层类脂膜包裹,脂膜内为基质蛋白(M1)层,膜上镶嵌着HA、NA和M2三种膜蛋白。HA和NA为流感病毒的主要抗原。根据HA和NA抗原性的差异,A型流感病毒可分16个HA亚型和9个NA亚型[1]。A型流感病毒具有广泛的宿主范围和超强的重组变异能力,对人类健康的威胁日趋…     

A型流感病毒RNA聚合酶及其对基因组复制和转录的调控作用

A型流感病毒是正粘病毒科成员,为单股负链分节段RNA病毒,全基因组由八个节段组成,分别编码八种结构蛋白(PB2、PB1、PA、HA、NP、NA、M1和M2)和两种非结构蛋白(NS1和NS2).核蛋白(NP)和RNA聚合酶复合体与病毒的八个RNA节段组成八个螺旋丝状的病毒核衣壳(RNP),核衣壳被双层类脂膜包裹,脂膜内为基质蛋白(M1)层,膜上镶嵌着HA、NA和M2三种膜蛋白.HA和NA为流感病毒的主要抗原.根据HA和NA抗原性的差异,A型流感病毒可分16个HA亚型和9个NA亚型[1].A型流感病毒具有广泛的宿主范围和超强的重组变异能力,对人类健康的威胁日趋严重,引起各国政府和科技工作者的广泛关注.研究RNA聚合酶的功能、揭示病毒复制和变异机理是目前抗流感病毒感染研究的热点之一.本文综述了流感病毒RNA聚合酶及其对病毒基因组复制和转录调控的研究进展.     

RNA病毒RNA聚合酶的研究进展

自然界仅有ＲＮＡ病毒以ＲＮＡ作为基因载体。依赖于ＲＮＡ的ＲＮＡ聚合酶在这种病毒的增殖复制期起到了非常重要的作用。它一方面以病毒ＲＮＡ为模板复制子代病毒的基因,另一方面也将病毒增殖期间需要的蛋白质和酶类的基因转录成为ｍＲＮＡ,也就是说它担负了复制酶和转...     

双链RNA病毒的复制机制

本文对双链ＲＮＡ（ｄｓＲＮＡ）病毒复制机制的研究进展进行了综述。根据ｄｓＲＮＡ病毒复制周期,分以下几方面对其复制的有关分子进行分析：转录、转录本释放和“满头”复制模型、（＋）ＲＮＡ转译、病毒包装、（－）ＲＮＡ合成等。此外,还简要分析了宿主基因及其产物在病毒复制中的作用。     

草鱼出血病病毒基因组体内转录的研究

本文采用α-[~(32)p]ATP标记物在鱼肾细胞系(CIK)系统中,对草鱼出血病病毒(Grass carp hemorrhage virus,GCHV)基因组进行了体内转录的研究。通过放线菌素D抑制宿主细胞基因组的转录活动,从感染病毒细胞中分离出病毒的mRNA,分别采用液相杂交和Nortbern blot方法检查病毒mRNA的转录情况。试验结果表明,GCHV含有内源性转录酶,其基因组的转录活动是在病毒感染细胞后4小时开始,由早期基因所转录,8—10小时获得晚期基因的转录产物。这些mRNA的大小、数目大体上与病毒基因组一致。     

植物双生病毒的复制及转录调控研究进展

双生病毒(Geminiviruses)是一类具有孪生颗粒形态的植物病毒,颗粒大小约为18nm×30nm.双生病毒成员众多,大致可分为三个亚组.双生病毒通常以粉虱或叶蝉为媒介进行传播,侵染范围十分广泛,对农业生产造成巨大的危害.感病植株一般表现为花叶、曲叶、黄化等症状,严重地影响了植物的正常生长.最近几年,属于双生病毒亚组Ⅲ的棉花曲叶病毒(Cotton leaf curl virus,CLCuV)在印巴次大陆广泛流行,严重时可使棉花绝收.双生病毒基因组DNA为单链环状形式,长度约为2 400nt～3 000nt.对双生病毒基因组结构、遗传表达机制,及其分类和进化进行深入的了解,有助于揭示寄主植物与病毒相互间的作用方式,进而为防治由双生病毒引起的植物病害奠定基础.本文就上述内容的最新进展作一简要综述.     

野田村病毒RNA的复制机制

野田村病毒科Nodaviradae分为2个属,分别为主要感染昆虫的α野田村病毒属(Alphanodavirus)和主要感染鱼类的β野田村病毒属(Betanodavirus)。野田村病毒的基因组由2条单链正义RNA分子(RNA1和RNA2)所组成,RNA1编码蛋白A,即病毒负责复制病毒两条基因组的依赖RNA的RNA聚合酶催化亚基。RNA2编码衣壳前体蛋白α,此前体蛋白α先组装成原病毒粒子,再经历一次自我催化的成熟切割成2个病毒的衣壳蛋白β和γ,就成了成熟的有感染性的病毒粒子。在RNA复制过程中,从RNA1的3′末端会合成一个不被包装进病毒粒子的亚基因组RNA3。RNA1能在无RNA2的情况下自我复制,并持续地产生亚基因组RNA3,RNA3的合成采取的是提前终止机制。本文还介绍了野田村病毒复制的调节、非结构蛋白的功能和病毒复制在细胞内的定位。     

单股环DNA病毒基因组的复制及其转录调控因子结合序列

单股环ＤＮＡ病毒基因组的复制及其转录调控因子结合序列崔治中（扬州大学农学院兽医系,生物技术系,扬州２２５００１）关键词单股环ＤＮＡ病毒,基因组复制,调控区结构最近定名的单股环ＤＮＡ病毒科（Ｃｉｒｃｏｖｉｒｉｄａｅ）是迄今为止发现的一类最小的动物病毒。...     

Molecular mechanisms of transcription and replication of the influenza A virus genome

Influenza A virus is one of the major pathogens that pose a large threat to human health worldwide and has caused pandemics.Influenza A virus is the Orthomyxoviridae prototype,and has 8 segmented negat...     

Assays for RNA synthesis and replication by the hepatitis C virus

At least six major genotypes of Hepatitis C virus (HCV) cause liver diseases worldwide.The efficacy rates with current standard of care are about 50％ against genotype 1,the most prevalent strain in the...     

Wrapping things up about virus RNA replication

All single-stranded 'positive-sense' RNA viruses that infect mammalian, insect or plant cells rearrange internal cellular membranes to provide an environment facilitating virus replication. A striking feature of these unique membrane structures is the induction of 70-100 nm vesicles (either free within the cytoplasm, associated with other induced vesicles or bound within a surrounding membrane) harbouring the viral replication complex (RC). Although similar in appearance, the cellular composition of these vesicles appears to vary for different viruses, implying different organelle origins for the intracellular sites of viral RNA replication. Genetic analysis has revealed that induction of these membrane structures can be attributed to a particular viral gene product, usually a non-structural protein. This review will highlight our current knowledge of the formation and composition of virus RCs and describe some of the similarities and differences in RNA-membrane interactions observed between the virus families Flaviviridae and Picornaviridae.     

RNA interference mediated inhibition of Chikungunya virus replication in mammalian cells

Chikungunya has emerged as one of the most important arboviral infection of public health significance. Recently several parts of Indian Ocean islands and India witnessed explosive, unprecedented epidemic. So far, there is no effective antiviral or licensed vaccine available against Chikungunya infection. RNA interference mediated inhibition of viral replication has emerged as a promising antiviral strategy. In this study, we examined the effectiveness of small interfering RNAs (siRNAs) against the inhibition of Chikungunya virus replication in Vero cells. Two siRNAs against the conserved regions of nsP3 and E1 genes of Chikungunya virus were designed. The siRNA activity was assessed by detecting both the infectious virus and its genome. The results indicated a reduction of virus titer up to 99.6% in siRNA transfected cells compared to control. The viral inhibition was most significant at 24 h (99%), followed by 48 h (65%) post infection. These results were also supported by the quantitative RT-PCR assay revealing similar reduction in Chikungunya viral genomic RNA. The siRNAs used had no effect on the expression of house keeping gene indicating non-interference in cellular mechanism. The specific and marked reduction in viral replication against rapidly replicating Chikungunya virus achieved in this study offers a potential new therapeutic approach. This is the first report demonstrating the effectiveness of siRNA against in vitro replication of Chikungunya virus.     

RNA binding protein 24 regulates the translation and replication of hepatitis C virus

The secondary structures of hepatitis C virus (HCV) RNA and the cellular proteins that bind to them are important for modulating both translation and RNA replication. However, the sets of RNA-binding proteins involved in the regulation of HCV translation, replication and encapsidation remain unknown. Here, we identified RNA binding motif protein 24 (RBM24) as a host factor participated in HCV translation and replication. Knockdown of RBM24 reduced HCV propagation in Huh7.5.1 cells. An enhanced translation and delayed RNA synthesis during the early phase of infection was observed in RBM24 silencing cells. However, both overexpression of RBM24 and recombinant human RBM24 protein suppressed HCV IRES-mediated translation. Further analysis revealed that the assembly of the 80S ribosome on the HCV IRES was interrupted by RBM24 protein through binding to the 5′-UTR. RBM24 could also interact with HCV Core and enhance the interaction of Core and 5′-UTR, which suppresses the expression of HCV. Moreover, RBM24 enhanced the interaction between the 5′- and 3′-UTRs in the HCV genome, which probably explained its requirement in HCV genome replication. Therefore, RBM24 is a novel host factor involved in HCV replication and may function at the switch from translation to replication.     

The first two nucleotides of the respiratory syncytial virus antigenome RNA replication product can be selected independently of the promoter terminus

There is limited knowledge regarding how the RNA-dependent RNA polymerases of the nonsegmented negative-strand RNA viruses initiate genome replication. In a previous study of respiratory syncytial virus (RSV) RNA replication, we found evidence that the polymerase could select the 5'-ATP residue of the genome RNA independently of the 3' nucleotide of the template. To investigate if a similar mechanism is used during antigenome synthesis, a study of initiation from the RSV leader (Le) promoter was performed using an intracellular minigenome assay in which RNA replication was restricted to a single step, so that the products examined were derived only from input mutant templates. Templates in which Le nucleotides 1U, or 1U and 2G, were deleted directed efficient replication, and in both cases, the replication products were initiated at the wild-type position, at position -1 or -2 relative to the template, respectively. Sequence analysis of the RNA products showed that they contained ATP and CTP at the -1 and -2 positions, respectively, thus restoring the mini-antigenome RNA to wild-type sequence. These data indicate that the RSV polymerase is able to select the first two nucleotides of the antigenome and initiate at the correct position, even if the 3'-terminal two nucleotides of the template are missing. Substitution of positions +1 and +2 of the template reduced RNA replication and resulted in increased initiation at positions +3 and +5. Together these data suggest a model for how the RSV polymerase initiates antigenome synthesis.