Institute for Clinical and Molecular Virology, Friedrich-Alexander University of Erlangen-Nürnberg (FAU), 91054 Erlangen, Germany.
Hannover Medical School, Institute of Virology, 30625 Hannover, Germany.
Viruses. 2020 Mar 11;12(3):303. doi: 10.3390/v12030303.
Nuclear egress is a regulated process shared by α-, β- and γ-herpesviruses. The core nuclear egress complex (NEC) is composed of the membrane-anchored protein homologs of human cytomegalovirus (HCMV) pUL50, murine cytomegalovirus (MCMV) pM50, Epstein-Barr virus (EBV) BFRF1 or varicella zoster virus (VZV) Orf24, which interact with the autologous NEC partners pUL53, pM53, BFLF2 or Orf27, respectively. Their recruitment of additional proteins leads to the assembly of a multicomponent NEC, coordinately regulating viral nucleocytoplasmic capsid egress. Here, the functionality of VZV, HCMV, MCMV and EBV core NECs was investigated by coimmunoprecipitation and confocal imaging analyses. Furthermore, a recombinant MCMV, harboring a replacement of ORF M50 by UL50, was analyzed both in vitro and in vivo. In essence, core NEC interactions were strictly limited to autologous NEC pairs and only included one measurable nonautologous interaction between the homologs of HCMV and MCMV. A comparative analysis of MCMV-WT versus MCMV-UL50-infected murine fibroblasts revealed almost identical phenotypes on the levels of protein and genomic replication kinetics. In infected BALB/c mice, virus spread to lung and other organs was found comparable between these viruses, thus stating functional complementarity. In conclusion, our study underlines that herpesviral core NEC proteins are functionally conserved regarding complementarity of core NEC interactions, which were found either virus-specific or restricted within subfamilies.
核出核是一种受调控的过程,α-、β-和 γ-疱疹病毒共有。核心核出核复合物(NEC)由人巨细胞病毒(HCMV)pUL50、鼠巨细胞病毒(MCMV)pM50、EBV BFRF1 或 VZV Orf24 的膜锚定蛋白同源物组成,它们分别与自身的 NEC 伴侣 pUL53、pM53、BFLF2 或 Orf27 相互作用。它们募集额外的蛋白质导致组装成一个多成分的 NEC,协调调节病毒核质衣壳出核。在这里,通过免疫共沉淀和共聚焦成像分析研究了 VZV、HCMV、MCMV 和 EBV 核心 NEC 的功能。此外,还分析了一种含有 ORF M50 被 UL50 取代的重组 MCMV,分别在体外和体内进行分析。本质上,核心 NEC 相互作用严格限于自身的 NEC 对,并且仅包括 HCMV 和 MCMV 同源物之间的一种可测量的非自身相互作用。对 MCMV-WT 与感染鼠成纤维细胞的 MCMV-UL50 的比较分析表明,在蛋白质和基因组复制动力学水平上几乎具有相同的表型。在感染的 BALB/c 小鼠中,发现这些病毒在肺部和其他器官中的病毒传播相当,因此具有功能互补性。总之,我们的研究强调了疱疹病毒核心 NEC 蛋白在核心 NEC 相互作用的互补性方面具有功能保守性,这些相互作用要么是病毒特异性的,要么局限于亚科内。