Genome sequence of a polydnavirus: insights into symbiotic virus evolution

Little is known of the fate of viruses involved in long-term obligatory associations with eukaryotes. For example, many species of parasitoid wasps have symbiotic viruses to manipulate host defenses and to allow development of parasitoid larvae. The complete nucleotide sequence of the DNA enclosed in the virus particles injected by a parasitoid wasp revealed a complex organization, resembling a eukaryote genomic region more than a viral genome. Although endocellular symbiont genomes have undergone a dramatic loss of genes, the evolution of symbiotic viruses appears to be characterized by extensive duplication of virulence genes coding for truncated versions of cellular proteins.     

Norwalk virus genome cloning and characterization

Major epidemic outbreaks of acute gastroenteritis result from infections with Norwalk or Norwalk-like viruses. Virus purified from stool specimens of volunteers experimentally infected with Norwalk virus was used to construct recombinant complementary DNA (cDNA) and derive clones representing most of the viral genome. The specificity of the clones was shown by their hybridization with post- (but not pre-) infection stool samples from volunteers infected with Norwalk virus and with purified Norwalk virus. A correlation was observed between the appearance of hybridization signals in stool samples and clinical symptoms of acute gastroenteritis in volunteers. Hybridization assays between overlapping clones, restriction enzyme analyses, and partial nucleotide sequence information of the clones indicated that Norwalk virus contains a single-stranded RNA genome of positive sense, with a polyadenylated tail at the 3' end and a size of at least 7.5 kilobases. A consensus amino acid sequence motif typical of viral RNA-dependent RNA polymerases was identified in one of the Norwalk virus clones. The availability of Norwalk-specific cDNA and the new sequence information of the viral genome should permit the development of sensitive diagnostic assays and studies of the molecular biology of the virus.     

Mapping of herpes simplex virus-1 neurovirulence to gamma 134.5, a gene nonessential for growth in culture

The gene designated gamma 134.5 maps in the inverted repeats flanking the long unique sequence of herpes simplex virus-1 (HSV-1) DNA, and therefore it is present in two copies per genome. This gene is not essential for viral growth in cell culture. Four recombinant viruses were genetically engineered to test the function of this gene. These were (i) a virus from which both copies of the gene were deleted, (ii) a virus containing a stop codon in both copies of the gene, (iii) a virus containing after the first codon an insert encoding a 16-amino acid epitope known to react with a specific monoclonal antibody, and (iv) a virus in which the deleted sequences were restored. The viruses from which the gene was deleted or which carried stop codons were avirulent on intracerebral inoculation of mice. The virus with the gene tagged by the sequence encoding the epitope was moderately virulent, whereas the restored virus reacquired the phenotype of the parent virus. Significant amounts of virus were recovered only from brains of animals inoculated with virulent viruses. Inasmuch as the product of the gamma 134.5 gene extended the host range of the virus by enabling it to replicate and destroy brain cells, it is a viral neurovirulence factor.     

Human immunodeficiency virus may encode a novel protein on the genomic DNA plus strand

The genome of the human immunodeficiency virus (HIV) is known to contain eight open reading frames (ORFs) on the minus strand of the double-stranded DNA replicative intermediate. Data presented here indicate that the DNA plus strand of HIV contains a previously unidentified ORF in a region complementary to the envelope gene sequence. This ORF could encode a protein of approximately 190 amino acid residues with a relative molecular mass of 20 kilodaltons if translation began from the first initiation codon. The predicted protein is highly hydrophobic and thus could be membrane associated. It is possible, therefore, that the HIV genome encodes a protein on antisense messenger RNA.     

A new class of endogenous human retroviral genomes

Human DNA contains multiple copies of a novel class of endogenous retroviral genomes. Analysis of a human recombinant DNA clone (HLM-2) containing one such proviral genome revealed that it is a mosaic of retroviral-related sequences with the organization and length of known endogenous retroviral genomes. The HLM-2 long terminal repeat hybridized with the long terminal repeat of the squirrel monkey virus, a type D retrovirus. The HLM-2 gag and pol genes share extensive nucleotide sequence homology with those of the M432 retrovirus (a type A-related retrovirus), mouse mammary tumor virus (a type B retrovirus), and the avian Rous sarcoma virus (a type C retrovirus). Nucleotide sequence analysis revealed regions in the HLM-2 pol gene that were as much as 70 percent identical to the mouse mammary tumor virus pol gene. A portion of the putative HLM-2 env gene hybridized with the corresponding region of the M432 viral genome.     

APOBEC-mediated editing of viral RNA

Retroviral DNA can be subjected to cytosine-to-uracil editing through the action of members of the APOBEC family of cytidine deaminases. Here we demonstrate that APOBEC-mediated cytidine deamination of human immunodeficiency virus (HIV) virion RNA can also occur. We speculate that the natural substrates of the APOBEC enzymes may extend to RNA viruses that do not replicate through DNA intermediates. Thus, cytosine-to-uracil editing may contribute to the sequence diversification of many viruses.     

Nucleotide sequence of yellow fever virus: implications for flavivirus gene expression and evolution

The sequence of the entire RNA genome of the type flavivirus, yellow fever virus, has been obtained. Inspection of this sequence reveals a single long open reading frame of 10,233 nucleotides, which could encode a polypeptide of 3411 amino acids. The structural proteins are found within the amino-terminal 780 residues of this polyprotein; the remainder of the open reading frame consists of nonstructural viral polypeptides. This genome organization implies that mature viral proteins are produced by posttranslational cleavage of a polyprotein precursor and has implications for flavivirus RNA replication and for the evolutionary relation of this virus family to other RNA viruses.     

种番鸭源禽坦布苏病毒分离鉴定及基因组序列分析

种番鸭是否感染禽坦布苏病毒颇受国内学者关注。本研究于国内外首次自表现产蛋下降的种番鸭卵巢中分离获得1株病毒（命名为WYZLJ-1359株）,经RT-PCR方法鉴定为禽坦布苏病毒。经比较分析该病毒与我国其他禽源分离株全基因组序列表明,种番鸭源分离株WYZLJ-1359主要分子特征未出现变异,与2010年以来我国分离的其他禽源分离毒株的核苷酸序列同源性均在98.4％以上,且亲缘关系非常近,遗传距离均在1.8％以下,远低于与2000年以前分离的坦布苏病毒分离株之间的遗传距离。以上结果表明,我国禽坦布苏病毒感染的宿主在不断增加,但病毒在不同品种家禽的传播过程中遗传较为稳定,基因组未出现基因的缺失或插入等变异现象。     

Extensive DNA inversions in the B. fragilis genome control variable gene expression

The obligately anaerobic bacterium Bacteroides fragilis, an opportunistic pathogen and inhabitant of the normal human colonic microbiota, exhibits considerable within-strain phase and antigenic variation of surface components. The complete genome sequence has revealed an unusual breadth (in number and in effect) of DNA inversion events that potentially control expression of many different components, including surface and secreted components, regulatory molecules, and restriction-modification proteins. Invertible promoters of two different types (12 group 1 and 11 group 2) were identified. One group has inversion crossover (fix) sites similar to the hix sites of Salmonella typhimurium. There are also four independent intergenic shufflons that potentially alter the expression and function of varied genes. The composition of the 10 different polysaccharide biosynthesis gene clusters identified (7 with associated invertible promoters) suggests a mechanism of synthesis similar to the O-antigen capsules of Escherichia coli.     

Amplification and molecular cloning of HTLV-I sequences from DNA of multiple sclerosis patients

Techniques of gene amplification, molecular cloning, and sequence analysis were used to test for the presence of sequences related to human T-lymphotropic virus type I (HTLV-I) in peripheral blood mononuclear cells of six patients with multiple sclerosis (MS) and 20 normal individuals. HTLV-I sequences were detected in all six MS patients and in one individual from the control group by DNA blot analysis and molecular cloning of amplified DNAs. The viral sequence in MS patients were associated with adherent cell populations consisting predominantly of monocytes and macrophages. Molecular cloning and nucleotide sequence analysis indicated that these amplified viral sequences were related to the HTLV-I proviral genome.     

茶蚕颗粒体病毒AcMNPVORF38同源基因的克隆及序列分析

[目的]为利用昆虫病毒更好地防治害虫和进行分子生物学研究。[方法]从感病茶蚕幼虫的尸体中提取颗粒体病毒DNA后,进行酶切、克隆、测序和序列分析。[结果]成功克隆了茶蚕颗粒体病毒基因组中一个1 484 bp的片段。该片段含有1个完整的开放阅读框(ORF)和2个不完整ORF。完整ORF为666 bp,有典型Nudix结构域,其编码1个与苜蓿银纹夜蛾核多角体病毒AcMNPV的ORF38高度同源的基因,定名为AcORF38同源基因。2个不完整ORF分别编码p47蛋白基因的C-端和p24基因的N-端。[结论]茶蚕颗粒体病毒与卷蛾科宿主中颗粒体病毒亲源关系较近,而与夜蛾科宿主昆虫中颗粒体病毒较远。AcORF38同源基因与编码NTP焦磷酸水解酶的基因高度同源,可能与病毒的复制和DNA损伤的修复过程相关,也可能与入侵寄主的过程相关。     

布尼亚病毒科全基因组序列比对分析*

 布尼亚科病毒（Bunyaviridae）是一类医学和农业上的重要病毒。本文从NCBI数据库下载具有完整基因组序列的布尼亚科病毒的5个属20种病毒的序列,用生物学软件DNAman,DNAstar进行比对分析,发现布尼亚科病毒属中的植物病毒在核酸序列及蛋白结构上与动物病毒有很大差异：（1）只有植物病毒的M基因组能编码NSm运动蛋白;（2）植物病毒和动物病毒在核酸序列和蛋白质序列长度均有差异,表明该科病毒是进化速度较快的病毒;（3）植物病毒核酸序列的GC含量低于动物病毒;（4）通过SMART网络软件进行蛋白质拓扑结构分析发现植物病毒和动物病毒在糖蛋白G_nG_c的结构上存在显著差异;（5）植物病毒糖蛋白结构较为复杂,有较多的紊乱区域,除INSV外,其他病毒都具有N端信号肽。这将对以后分子生物学检测中引物的设计及病毒的鉴定等方面的研究起到积极的作用。     

The Genome sequence of the SARS-associated coronavirus

We sequenced the 29,751-base genome of the severe acute respiratory syndrome (SARS)-associated coronavirus known as the Tor2 isolate. The genome sequence reveals that this coronavirus is only moderately related to other known coronaviruses, including two human coronaviruses, HCoV-OC43 and HCoV-229E. Phylogenetic analysis of the predicted viral proteins indicates that the virus does not closely resemble any of the three previously known groups of coronaviruses. The genome sequence will aid in the diagnosis of SARS virus infection in humans and potential animal hosts (using polymerase chain reaction and immunological tests), in the development of antivirals (including neutralizing antibodies), and in the identification of putative epitopes for vaccine development.     

烟草坏死病毒完整基因组的统计特征与聚类

[目的]提取烟草坏死病毒完整基因组的统计特征。并且对其进行聚类分析。[方法]在烟草坏死病毒完整基因组的碱基序列上,用每个碱基及其随后2个碱基所构成的三碱基组排列成一个新的序列S;计算所有64种不同三碱基组在S上出现的概率,得到一个64维向量L;比较各个基因组的L向量,得到8个三碱基组,它们的概率存在明显差异。[结果]8个三碱基组（CGT;CAA;TAC;TAA;AGG;ATG;ATA;AAG）的出现概率与烟草坏死病毒基因组的遗传变异有着重要关联;4个不同来源的烟草坏死病毒完整基因组,按其遗传变异结果形成2个大类。[结论]该研究方法普遍适用于各种烟草病毒基因序列的分析,为烟草坏死病毒病的防治提供理论依据。     

烟草坏死病毒完整基因组的统计特征与聚类（摘要）（英文）

[目的]提取烟草坏死病毒完整基因组的统计特征,并对其进行聚类分析。[方法]在烟草坏死病毒完整基因组的碱基序列上,用每个碱基及其随后2个碱基所构成的三碱基组排列成一个新的序列S;计算所有64种不同三碱基组在S上出现的概率,得到一个64维向量L;比较各个基因组的L向量,得到8个三碱基组,它们的概率存在明显差异;依照一定的规律,给三碱基组赋予数字代码;最后,对不同来源的烟草坏死病毒完整基因组按照L向量进行K-M聚类分析。[结果]8个三碱基组（CGT;CAA;TAC;TAA;AGG;ATG;ATA;AAG）的出现概率与烟草坏死病毒基因组的遗传变异有着重要关联;4个不同来源的烟草坏死病毒完整基因组,按其遗传变异结果形成2个大类。[结论]该研究方法普遍适用于各种烟草病毒基因序列的分析,为烟草坏死病毒病的防治提供了理论依据。     

Homology of cytokine synthesis inhibitory factor (IL-10) to the Epstein-Barr virus gene BCRFI

Complementary DNA clones encoding mouse cytokine synthesis inhibitory factor (CSIF; interleukin-10), which inhibits cytokine synthesis by TH1 helper T cells, were isolated and expressed. The predicted protein sequence shows extensive homology with an uncharacterized open reading frame, BCRFI, in the Epstein-Barr virus genome, suggesting the possibility that this herpes virus exploits the biological activity of a captured cytokine gene to enhance its survival in the host.