Human metapneumovirus nucleoprotein and phosphoprotein interact and provide the minimal requirements for inclusion body formation.
Human metapneumovirus nucleoprotein and phosphoprotein interact and provide the minimal requirements for inclusion body formation.
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DOI：
10.1099/vir.0.2008/004051-0
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发表时间：
2008-11
期刊：
影响因子：
--
通讯作者：
Crowe JE
中科院分区：
文献类型：
--
作者：
Derdowski A;Peters TR;Glover N;Qian R;Utley TJ;Burnett A;Williams JV;Spearman P;Crowe JE
人偏肺病毒(HMPV)是新近发现的一种新发现的肺炎病毒亚科副粘病毒，还包括禽肺炎病毒和人呼吸道合胞病毒(HRSV)。HMPV是世界范围内呼吸道疾病的重要原因。为了了解HMPV复制的早期事件，从感染A1型HMPV野毒株TN/96-12的细胞中克隆了编码HMPV核蛋白(N)、磷蛋白(P)、基质蛋白(M)、M2-1蛋白和M2-2蛋白的cDNA。HMPV N和P通过多种技术相互作用：酵母双杂交、免疫共沉淀和荧光共振能量转移(FRET)。共聚焦显微镜研究表明，单独表达时，荧光标记的HMPV N和P在未感染细胞的宿主细胞质中呈弥漫性表达，但在感染HMPV的细胞中被募集到细胞质病毒包涵体中。此外，当HMPV N和P一起表达时，即使在没有病毒感染的情况下，它们也形成了细胞质内包涵体样复合体。FRET显微镜显示HMPV N和P在细胞质包涵体内直接相互作用。此外，酵母双杂交分析表明，N-末端28个氨基酸是募集和形成细胞质内含体所必需的，但对于与HMPV P结合是必不可少的。这表明HMPV N和P蛋白为HMPV包涵体的形成提供了最低限度的病毒需求，这可能是肺炎病毒亚科成员的一个区别特征。
Human metapneumovirus (HMPV) is a recently discovered paramyxovirus of the subfamily Pneumovirinae, which also includes avian pneumovirus and human respiratory syncytial virus (HRSV). HMPV is an important cause of respiratory disease worldwide. To understand early events in HMPV replication, cDNAs encoding the HMPV nucleoprotein (N), phosphoprotein (P), matrix protein (M), M2-1 protein and M2-2 protein were cloned from cells infected with the genotype A1 HMPV wild-type strain TN/96-12. HMPV N and P were shown to interact using a variety of techniques: yeast two-hybrid assays, co-immunoprecipitation and fluorescence resonance energy transfer (FRET). Confocal microscopy studies showed that, when expressed individually, fluorescently tagged HMPV N and P exhibited a diffuse expression pattern in the host-cell cytoplasm of uninfected cells but were recruited to cytoplasmic viral inclusion bodies in HMPV-infected cells. Furthermore, when HMPV N and P were expressed together, they also formed cytoplasmic inclusion-like complexes, even in the absence of viral infection. FRET microscopy revealed that HMPV N and P interacted directly within cytoplasmic inclusion-like complexes. Moreover, it was shown by yeast two-hybrid analysis that the N-terminal 28 aa are required for the recruitment to and formation of cytoplasmic inclusions, but are dispensable for binding to HMPV P. This work showed that HMPV N and P proteins provide the minimal viral requirements for HMPV inclusion body formation, which may be a distinguishing characteristic of members of the subfamily Pneumovirinae.
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影响因子：
5.4
作者：
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