乌库尼米病毒核衣壳蛋白正电荷氨基酸残基的突变分析。

Mutational analysis of positively charged amino acid residues of Uukuniemi phlebovirus nucleocapsid protein.

机构信息

Research Programs Unit, Infection Biology Research Program, Department of Virology, Haartman Institute, PO Box 21, University of Helsinki, 00014 Helsinki, Finland.

出版信息

Virus Res. 2012 Jul;167(1):118-23. doi: 10.1016/j.virusres.2012.04.003.

DOI:10.1016/j.virusres.2012.04.003

PMID:22808531

Abstract

The aim of this study was to evaluate the contribution of positively charged amino acid residues for the Uukuniemi virus (UUKV) N protein functionality. Based on phlebovirus nucleocapsid (N) protein alignments and 3D-structure predictions of UUKV N protein, 14 positively charged residues were chosen as targets for the mutagenesis. The impact of mutations to the N protein functionality was analyzed using minigenome-, virus-like particle-, and mammalian two-hybrid-assays. Seven of the mutations affected the functional competence in all three assays, while others had milder impact or no impact at all. In the 3D-model of UUKV N protein, five of the affected residues, R61, R64, R73, R98 and R115, were located either within or in close proximity to the central cavity that could potentially bind RNA.

摘要

本研究旨在评估正电荷氨基酸残基对 Uukuniemi 病毒（UUKV）N 蛋白功能的贡献。基于黄病毒核衣壳（N）蛋白的比对和 UUKV N 蛋白的 3D 结构预测，选择了 14 个正电荷残基作为突变靶点。使用小基因、病毒样颗粒和哺乳动物双杂交测定分析突变对 N 蛋白功能的影响。其中 7 个突变在所有三种测定中均影响功能，而其他突变的影响则较轻或根本没有影响。在 UUKV N 蛋白的 3D 模型中，受影响的 5 个残基 R61、R64、R73、R98 和 R115 位于或靠近中央腔，该腔可能结合 RNA。