Molecular mechanism on the replication of Bombyx densovirus

家蚕浓核病毒复制的分子机制

• 批准号: 07456032
• 负责人: BANDO Hisanori
• 金额: $ 4.99万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07456032/
• 关键词: DNA VIRUS; HOST SPECIFICITY; REGULATORY PROTEIN; TRANSACTIVATOR; GENOME REPLICATION; 昆虫ウイルス; デンソウイルス; 遺伝子複製機構

Bombyx mori denso nucleosis virus (Yamanashi isolate ; BmDNV-2) is a small spherical virus with a bi-partite genome. In this study, to elucidate the nature of BmDNV-2, the viral DNA replication mechanism, genome organization and function of viral proteins were investigated. Analysis of the replicative intermediates (RI) of the viral DNA suggested the self-priming and hairpin-transfer model for parvovirus replication could not be applied to the replication of BmDNV-2. Nucleotide sequencing studies of the genomic DNAs showed that, unlike BmDNV-1 and the other parvoviruses the BmDNV-2 terminal sequences did not contained the large palindromic sequence which could form a stable hairpin structure. This was confirmed by the secondary structure prediction using a computer analyzes program. These results also suggested that the BmDNV-2 had a unique replication mechanisms other than the self-priming and hairpin-transfer mechanism.Furthermore, the sequence analyzes elucidated that the BmDNV-2 ge … More nome contained 6 major open reading frames (ORFs), 4 ORFs in VD1 and 2in VD2. Immunochemical studies demonstrated that the major four viral structural proteins (VP1,2,3 and 4) were encoded in the ORF2 of VD1. Followingly, the functions of the nonstructural proteins (NSPs) were investigated by transient expression assay using the luciferase as a reporter. The NSPs, p37 and p14 which were encoded in ORF3 and4 of VD1, respectively, transactivated the tentative promoter sequence for the major structural protein gene, suggesting that these proteins were concerned in the regulation of the expression fo structural proteins. In addition, p27 which was an NSP encoded in VD2 could transactivate all of the viral promoters implying that p27 is a key regulatory protein in the BmDNV-2 replication.In conclusion, this study elucidated the genome structure and the genome organization of the BmDNV-2. The results revealed that the BmDNV-2 is a new type of single-stranded DNA virus with a unique replication mechanism which was different from those of parvoviruses. Furthermore, the functional analyzes of the viral proteins suggested that the viral gene expression was regulated in the complicated manner by the proteins encoded in viral DNAs. And, p27 will be a key regulatory protein in the replication of the BmDNV-2 and may be one of the factor concerning in the determination of the host specificity. Less

家蚕浓核病毒（Yamanashi isolate ; BmDNV-2）是具有二分基因组的小型球形病毒。本研究对BmDNV-2的DNA复制机制、基因组结构和病毒蛋白的功能进行了研究，旨在阐明BmDNV-2的本质。对病毒DNA复制中间体的分析表明，细小病毒复制的自启动和发夹转移模型不能应用于BmDNV-2的复制。基因组DNA的核苷酸测序研究表明，与BmDNV-1和其他细小病毒不同，BmDNV-2的末端序列不包含可以形成稳定发夹结构的大回文序列。这通过使用计算机分析程序的二级结构预测得到证实。这些结果也表明BmDNV-2除了自身启动和发夹转移机制外，还具有独特的复制机制，序列分析进一步证实了BmDNV-2基因是一种具有生物学活性的基因。 ...更多信息 NOME包含6个主要的开放阅读框（ORF），其中VD 1有4个，VD 2有2个。免疫组化研究表明，VD 1的ORF 2编码4种主要的病毒结构蛋白（VP 1，2，3和4）。随后，利用荧光素酶作为报告基因，通过瞬时表达分析研究了非结构蛋白（NSP）的功能。VD 1的ORF 3和ORF 4编码的NSPs p37和p14分别反式激活了主要结构蛋白基因的启动子序列，表明这些蛋白参与了结构蛋白的表达调控。另外，VD 2编码的NSP p27可以反式激活所有的病毒启动子，提示p27是BmDNV-2复制的关键调控蛋白。结果表明，BmDNV-2是一种新型的单链DNA病毒，具有不同于细小病毒的独特复制机制。此外，对病毒蛋白的功能分析表明，病毒基因的表达受到病毒DNA编码蛋白的复杂调控。p27可能是BmDNV-2复制过程中的一个关键调控蛋白，也可能是决定宿主特异性的因素之一。少

项目成果

Tohru Hayakawa: "Detection of replicative intermechiate with closed terminus of donsonucleosis virus" Archives of Virology. 142. 1-7 (1997)

Tohru Hayakawa：“检测具有Donsonucleosis病毒封闭末端的复制中间体”病毒学档案。

Tohru Hayakawa: "Detection of replicative intermediate with closed terminus of Bombyx densonucleosis virus" Archives of Virology. 142. 1-7 (1997)

Tohru Hayakawa：“检测具有家蚕浓核病毒封闭末端的复制中间体”病毒学档案。

Hayakawa Tohru, Asano Shin-ichiro, Sahara Ken, Iizuka Toshihiko, and Bando Hisanori: "Detection of replicative intermediate with closed terminus of Bombyx densonucleosis virus." Arch.Virol.142. 1-7 (1997)

Hayakawa Tohru、Asano Shin-ichiro、Sahara Ken、Iizuka Toshihiko 和 Bando Hisanori：“检测具有 Bombyx densonucleosis 病毒封闭末端的复制中间体。”

Bando, H., T.Hayakawa, S.Asano, K,Sahara, M.Nakagaki, and T.Iizuka: "Analysis on the genetic information in a DNA segment of a virus of new type from silkworm." Arch.Virol.140. 1147-1155 (1995)

Bando, H.、T.Hayakawa、S.Asano、K,Sahara、M.Nakagaki 和 T.Iizuka：“蚕新型病毒 DNA 片段的遗传信息分析”。

Hisanori Bando: "Analysis on the genetic imtormation in a DNA segment of a winus of new type from silkworm" Archives of Virology. 140. 1147-1155 (1995)

Hisanori Bando：“来自蚕的新型 Winus 的 DNA 片段的遗传信息分析”病毒学档案。