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胡远扬, Xiaochun Wang, Jiamin Zhang, Jie Lu, Fuming Yi, Chuanfeng Liu and Yuanyang Hu
Journal of General Virology (2004), 85, 1145-1151,-0001,():
-1年11月30日
The complete nucleotide sequence of a new insect picorna-like virus, Ectropis obliqua picorna-like virus (EoPV), which causes a fatal infection of Ectropis obliqua larvae, has been determined. The genomic RNA of EoPV is 9394 nt in length and contains a single, large open reading frame (nt 391-9351) encoding a polyprotein of 2987 aa. Sequence comparisons with other viral polyproteins revealed that the consensus sequences for picornavirus RNA helicase, protease and RNA-dependent RNA polymerase proteins are found on the genome in order in the 59R39 direction. All structural genes were located at the 59 terminus. In terms of sequence similarity, identity and genome organization, EoPV resembles mammalian picornaviruses and three other insect picorna-like viruses: Infectious flacherie virus of silkworm, Sacbrood virus of honeybee and Perina nuda picorna-like virus (PnPV). Phylogenetic analysis showed that EoPV is most closely related to PnPV and suggests that these four insect picorna-like viruses might constitute a new group of insect-infectious RNA viruses.
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【期刊论文】Biochemical characterization of Periplaneta fuliginosa densovirus non-structural protein NS1
胡远扬, Bo Yang, Jiamin Zhang, Dawei Cai, Doulin Li, Wuguo Chen, Hong Jiang, Yuanyang Hu*
Biochemical and Biophysical Research Communications 342(2006)1188-1196,-0001,():
-1年11月30日
The non-structural (NS) proteins of parvoviruses are involved in essential steps of the viral life cycle. Various biochemical functions, such as ATP binding, ATPase, site-specific DNA binding and nicking, and helicase activities, have been assigned to the protein NS1. Compared with the non-structural proteins of the vertebrate parvoviruses, the NS proteins of the Densovirinae have not been well characterized. Here, we describe the biochemical properties of NS1 of Periplaneta fuliginosa densovirus (PfDNV). We have expressed and purified NS1 using a baculovirus system and analyzed its enzymatic activity. The purified recombinant NS1 protein possesses ATPaseand ATP-or dATP-dependent helicase activity requiring either Mg2+ or Mn2+ as a cofactor. The ATPase activity of NS1 can be efficiently stimulated by single-stranded DNA. The ATPase coupled helicase activity was detected on blunt-ended double-stranded oligonucleotide substrate. Using South-Western and Dot-spot assays, we identified a DNA fragment that is recognized specifically by the recombinant NS1 protein. The fragment consists of (CAC)4 and is located on the hairpin region of the terminal palindrome. The domain for DNA binding was defined to the amino-terminal region (amino acids 1-250). In addition, we found that NS1 can form oligomeric complexes in vivo and in vitro. Mutagenesis analysis showed that ATP binding is necessary for oligomerization. Based on these results, it seems that PfDNV NS1, a multifunctional protein, plays an important role in viral DNA replication comparable to those of vertebrate parvovirus initiator proteins.
Periplaneta fuliginosa densovirus, NS1, ATPase, Helicase, South-Western, Yeast two-hybrid, Oligomerization, Far-Western
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胡远扬, Fuming Yi, Jiamin Zhang, Haiyang Yu, Chuanfeng Liu, Junping Wang and Yuanyang Hu
Journal of General Virology (2005), 86, 789-796,-0001,():
-1年11月30日
In this study, Dendrolimus punctatus tetravirus (DpTV) has been identified as a new member of the genus Omegatetravirus of the family Tetraviridae that may be related serologically to Nudaurelia capensis v virus (NvV). DpTV particles are isometric, with a diameter of about 40nm and a buoyant density of 1?281 g cm3 in CsCl. The virus has two capsid proteins (of 62 500 and 6800 Da) and two single-stranded RNA molecules (RNA1 and RNA2), which are 5492 and 2490 nt long, respectively. RNA1 has a large open reading frame (ORF) encoding a polypeptide of 180 kDa; RNA2 contains two partially overlapping ORFs encoding polypeptides of 17 and 70 kDa. The 180 kDa protein, which contains consensus motifs of a putative methyltransferase, helicase and RNA-dependent RNA polymerase, shows significant similarity to those of other tetraviruses. The 17 kDa protein is a PEST (Pro/Glu/Ser/Thr) protein of unknown function. The 70 kDa protein is the coat protein precursor and is predicted to be cleaved at an Asn-Phe site located after residue 570. The 70 kDa protein shows 86 and 66% identity to its homologues in NvV and Helicoverpa armigera stunt virus, respectively. Secondary-structure analysis revealed that the RNAs of DpTV have tRNA-like structures at their 39 termini.
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胡远扬, 易福明, 张珈敏, 刘传凤, 胡远扬*
昆虫学报Acta Entomologica Sinica, October 2005, 48 (5): 792-798,-0001,():
-1年11月30日
T4病毒科由一类单股正链RNA病毒组成,分为松天蛾B样病毒属和松天蛾 样病毒属。这2个属的病毒具有不同的基因组结构,8样病毒含单组分基因组,其结构蛋白由一亚基因组RNA表达;而 样病毒含双组分基因组,2个RNA分子分别编码复制酶蛋白和结构蛋白。在T4病毒基因组RNA3 端有类似tRNA的二级结构。样病毒壳蛋白的氨基酸序列一致性高达66%~86&,而G样病毒壳蛋白的氨基酸同源性则要低得多。在昆虫细胞中表达壳蛋白基因时都能形成病毒类似粒子。该文还介绍了T4病毒复制机理以及T4病毒与其他病毒的进化关系。
昆虫, T4病毒, 基因组结构, tRNA类似结构, 系统发育
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【期刊论文】茶尺蠖小RNA病毒5'端非编码区的克隆和测序及与哺乳动物小RNA病毒的比较分析
胡远扬, 王小纯..张珈敏, 蒋洪, 俞海洋, 谭莉, 胡远扬*
昆虫学报Acta Entomologica Sinica, October 2004, 47 (5): 573-578,-0001,():
-1年11月30日
用Trizol从纯化的茶尺蠖Ectropis oblique小RNA病毒(EoPV)中提取病毒基因组RNA,逆转录后加poly(dT),然后进行两步PCR扩增基因组5'端。克隆测序后,对其5 端非编码区的核苷酸序列进行分析,发现具有哺乳动物小RNA病毒的5'端非编码区的一些特征:A/T含量丰富、起始密码子上游AUG和小顺反子多。利用mfold预测了EoPV5'端非编码区的二级结构,存在4个茎环结构,有哺乳动物内部核糖体进入位点(IRES)的保守区域,即含保守基序GNRA的茎环A和A/C丰富的环B及多聚嘧啶区域。据此推测EoPV基因组翻译采用IRES起始机制。
尺蠖小RNA病毒, 哺乳动物小RNA病毒, 5', 端非编码区, 结构分析
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