CCAAT/enhancer binding proteins play a role in oriLyt-dependent genome replication during MHV-68 de novo infection

CCAAT/enhancer binding proteins play a role in oriLyt-dependent genome replication during MHV-68 de novo infection
复制标题

CCAAT/增强子结合蛋白在 MHV-68 从头感染期间 oriLyt 依赖性基因组复制中发挥作用

DOI:
10.1007/s13238-011-1060-z
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发表时间:
2011-06
期刊:
Protein Cell
影响因子:
--
通讯作者:
Hongyu Deng
Hongyu Deng
中科院分区:
其他
文献类型:
--
作者:
Jing Qi;Danyang Gong;Hongyu Deng

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小鼠γ疱疹病毒68 (MHV-68)是γ疱疹病毒家族的一员,在受纳细胞系中大量复制,能够感染实验室小鼠。MHV-68已成为研究病毒复制的基本方面和其人类对应物的宿主-病毒相互作用的模型。疱疹病毒基因组的复制是通过病毒基因组中称为裂解复制起始(oriLyt)的顺式元件介导的。转录因子家族,CCAAT/增强子结合蛋白(C/ ebp),在γ疱疹病毒再激活过程中协助orilyl介导的DNA复制。在这项研究中,我们以MHV-68为模型,研究了C/ ebp在新发感染期间γ疱疹病毒DNA复制中的作用。在体外和体内,我们发现C/EBP α和β结合在MHV-68 oriLyt核心区域的CCAAT盒子上,电泳迁移率转移实验和染色质免疫沉淀实验证实了这一点。在复制实验中,C/ ebp的显性阴性形式显著降低了MHV-68在质粒和基因组水平上的裂解复制效率,表明功能性C/ ebp是最大限度地复制MHV-68基因组DNA所必需的。总的来说,我们的数据表明C/ ebp与oriLyt核心区域相互作用,并在新感染期间MHV-68裂解DNA复制中发挥重要作用。
Murine gammaherpesvirus 68 (MHV-68), a member of the gammaherpesvirus family, replicates robustly in permissive cell lines and is able to infect laboratory mice. MHV-68 has emerged as a model for studying the basic aspects of viral replication and host-virus interactions of its human counterparts. Herpesvirus genome replication is mediated through a cis-element in the viral genome called the origin of lytic replication (oriLyt). A family of transcription factors, CCAAT/enhancer binding proteins (C/EBPs), assists in oriLyt-mediated DNA replication during gammaherpesvirus reactivation. In this study, we examined the role of C/EBPs in gammaherpesvirus DNA replication during de novo infection, using MHV-68 as a model. We found that C/EBP α and β bind to the CCAAT boxes in the MHV-68 oriLyt core region both in vitro and in vivo, as demonstrated by electrophoretic mobility shift assay and chromatin immunoprecipitation assay. A dominant negative form of C/EBPs significantly impaired the lytic replication efficiency of MHV-68 on both the plasmid and genome levels in a replication assay, indicating that functional C/EBPs are required for maximal MHV-68 genome DNA replication. Collectively, our data demonstrate that C/EBPs interact with the oriLyt core region and play an important role in MHV-68 lytic DNA replication during de novo infection.
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