Identification of a human respiratory syncytial virus phosphoprotein domain required for virus-like-particle formation.
Identification of a human respiratory syncytial virus phosphoprotein domain required for virus-like-particle formation.
复制标题
病毒样颗粒形成所需的人呼吸道合胞病毒磷蛋白结构域的鉴定。
DOI：
10.1016/j.virol.2019.04.001
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发表时间：
2019
期刊：
影响因子：
3.7
通讯作者：
Oomens,AntoniusGP
中科院分区：
文献类型：
--
作者：
Meshram,ChetanD;Oomens,AntoniusGP
对人呼吸道合胞病毒（hRSV）组装过程的感知的低效率和不充分的知识对用于疫苗目的的真实hRSV病毒样颗粒（VLP）的大规模生产提出了障碍。我们先前确定基质蛋白、磷蛋白（P）和融合蛋白羧基末端足以产生类似于丝状野生型hRSV的VLP。在这里，研究了P的贡献。通过共表达基质、融合蛋白和修饰的P蛋白，发现富含Ser/Thr的P区域（残基39-57）对于VLP形成是关键的，而寡聚化结构域不是。整个区域39-57的取代抑制VLP形成，并鉴定了相关氨基酸。丝氨酸和苏氨酸的拟磷酸化取代抑制VLP形成;磷酸化取代没有。数据显示，P不仅共调节复制和转录，而且在组装中也具有重要作用，由可能与M和/或F相互作用的单独结构域介导，并且由磷酸化高度调节。
Perceived inefficiency and inadequate knowledge of the human respiratory syncytial virus (hRSV) assembly process present a hurdle for large-scale production of authentic hRSV virus-like particles (VLPs) for vaccine purposes. We previously established that the matrix protein, phosphoprotein (P), and fusion protein carboxy-terminus were sufficient to generate VLPs that resemble filamentous wildtype hRSV. Here, the contribution of P was examined. By co-expressing matrix, fusion, and modified P proteins, a ser/thr-rich P region (residues 39–57) was found to be critical for VLP formation, whereas the oligomerization domain was not. Substitutions throughout region 39–57 inhibited VLP formation and relevant amino acids were identified. Phosphomimetic substitutions of serines and threonines inhibited VLP formation; Phosphoblatant substitutions did not. The data show that P not only co-regulates replication and transcription but also has an important role in assembly, mediated by a separate domain that likely interacts with M and/or F and is highly regulated by phosphorylation.