Analysis and the latency of herpes simplex virus

单纯疱疹病毒的潜伏期分析

• 批准号: 11694254
• 负责人: SHIRAKATA Masaki
• 金额: $ 3.14万
• 依托单位: Tokyo Medical and Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11694254/
• 关键词: HSV; ICP0; latency; BMAL1; UL13; EF-1δ; 潜伏感染; 再活性化

The aim of the research was to investigate the mechanism of herpes simplex virus (HSV) latency. The results obtained during these two years were mainly as follows.(i) ICP0 is an important regulatory protein that is involved in regulation both of gene expression in productive infection and latency. None of the known functions at the molecular level account for the apparent activity of ICP0 as a transactivator. Here we report that ICP0 functionally interacts with a cellular transcription factor BMAL1, a member of the bHLH-PAS super family of transcriptional regulators. Specifically, sequences mapped to the exon II of ICP0 interacted with BMAL1 in the yeast two-hybrid system and in reciprocal pull down experiments in vitro.Moreover, the enhancement of transcription of a luciferase reporter construct whose promoter contained multiple BMAL1 binding sites by ICP0 and BMAL1 was significantly greater than that observed by ICP0 or BMAL1 alone.These results indicate that ICP0 interacts physicall … More y and functionally with at least one cellular transcription-regulatory factor. (ii) An earlier report (Y.Kawaguchi, R.Bruni, and B.Roizman, J.Virol. 71 : 1019-1024,1997 ; Y.Kawaguchi, C.Van Sant, and B.Roizman, J.Virol. 72 : 1731-1736,1998) showed that herpes simplex virus 1 (HSV-1) infection causes the hyperphosphorylation of translation elongation factor 1δ (EF-1δ) and that the modification of EF-1δ is the consequence of direct phosphorylation by a viral protein Kinase encoded by the U_L13 gene of HSV-1. The U_L13 gene is conserved in members of all herpesvirus subfamilies. Here we report the following. (i) In various mammalian cells, accumulation of the hyperphosphorylated form of EF-1δ is observed after infection with alpha-, beta- and gammaherpesviruses including HSV-2, feline herpesvirus type 1, pseudorabisvirus, and bovine herpesvirus type 1, human cytomegalovirus (HCMV) and equine herpesvirus type 2. (ii) In HLF cells infected with recombinant HSV-1 lacking U_L13 gene, the hypophosphorylated form of EF-1δ is a minor species, whereas the relative amount of the hyperphosphorylated form of EF-1δ significantly increases in cells infected with the recombinant HSV-1 in which U_L13 is replaced by HCMV U_L97, a homologue of U_L13. These results indicate that the post-translational modification of EF-1δ is conserved herpesvirus function and the U_L13 homologues may be responsible for the universal-modification of the translation factor. Less

本研究旨在探讨单纯疱疹病毒(HSV)潜伏的机制。主要研究结果如下：(1)ICP0是一种重要的调控蛋白，参与了产生性感染和潜伏期基因表达的调控。在分子水平上的任何已知功能都不能解释ICP0作为反式激活剂的明显活性。在这里，我们报告了ICP0与细胞转录因子BMAL1在功能上的相互作用，BMAL1是bHLH-PAS超家族转录调控因子的成员。在酵母双杂交系统和体外双向下拉实验中，定位于ICP0外显子II的序列与BMAL1相互作用。此外，ICP0和BMAL1对启动子含有多个BMAL1结合位点的荧光素酶报告结构的转录促进作用明显大于ICP0或BMAL1单独作用。这些结果表明，ICP0与物理…相互作用与至少一个细胞转录调节因子具有更多的y和功能。(Ii)较早的报告(Y.Kawaguchi，R.Bruni和B.Roizman，J.Virol)。71：1019-1024,1997；Y.Kawaguchi，C.Van Sant和B.Roizman，J.Virol。(72：1731-1736,1998)的研究表明，单纯疱疹病毒1型感染引起翻译延伸因子1δ(EF-1δ)的过度磷酸化，EF-1δ的修饰是单纯疱疹病毒U_L13基因编码的病毒蛋白激酶直接磷酸化的结果。U_L13基因在疱疹病毒各亚科成员中都是保守的。在这里，我们报道以下内容。(I)在多种哺乳动物细胞中，感染包括单纯疱疹病毒2型、猫疱疹病毒1型、伪狂犬病病毒、牛疱疹病毒1型、人巨细胞病毒和马疱疹病毒2型在内的重组单纯疱疹病毒后，在多种哺乳动物细胞中可以观察到高磷酸化的EF-1δ的积累。(Ii)在缺乏U_L13基因的重组单纯疱疹病毒感染的细胞中，低磷酸化的EF-1δ是次要的，而在由U_L13的同源物HCMV U_L97取代U_L13的重组HSV-1感染的细胞中，高磷酸化的EF-1δ的相对含量显著增加。这些结果表明，EF-1δ的翻译后修饰是保守的疱疹病毒功能，U_L13同源物可能是翻译因子普遍修饰的原因。较少

项目成果

Advani SJ, Weichselbaum RR, Roizman B.: "The role of cdc2 in the expression of herpes simplex virus genes."Proc.Natl Acad Sci U S A. 97(20). 10996-1001 (2000)

Advani SJ、Weichselbaum RR、Roizman B.：“cdc2 在单纯疱疹病毒基因表达中的作用。”Proc.Natl Acad Sci U S A. 97(20)。

Randall G, Lagunoff M, Roizman B.: "Herpes simplex virus 1 open reading frames O and P are not necessary for establishment of latent infection in mice."J Virol.. 74(19). 9019-27 (2000)

Randall G、Lagunoff M、Roizman B.：“单纯疱疹病毒 1 开放阅读框 O 和 P 对于在小鼠中建立潜伏感染不是必需的。”J Virol.. 74(19)。

Masaki Shirakata, Ken-Ichi Imadome, and Kanji Hirai.: "Requirement of replication licensing for the dyad symmetry element-dependent replication of the EBV oriP minichromosome."Virology. 263. 42-54 (1999)

Masaki Shirakata、Ken-Ichi Imadome 和 Kanji Hirai.：“EBV oriP 微型染色体的二元对称元件依赖性复制的复制许可要求。”病毒学。

Van Sant C et al.,: "Role of Cyclin D3 in the Biology of Herpes Simplex Virus 1 ICP0."J Virol.. 75. 1888-1898 (2001)

Van Sant C 等人，：“细胞周期蛋白 D3 在单纯疱疹病毒 1 ICP0 生物学中的作用。”J Virol.. 75. 1888-1898 (2001)

Y.Kawaguchi, T.Matsumuta, B.Roizman, and K.Hirai: "The cellular elongation factor 1δ is modified in cells infected with representative alpha-, beta, or gamma herpesviruses."J.Virol.. 73. 4456-4460 (1999)

Y.Kawaguchi、T.Matsumuta、B.Roizman 和 K.Hirai：“细胞伸长因子 1δ 在感染了代表性 α、β 或 γ 疱疹病毒的细胞中被修饰。”J.Virol.. 73. 4456-4460 (1999)