Watanabe N, Kawano M, Tsurudome M, Nishio M, Ito M, Ohgimoto S, Suga S, Komada H, Ito Y
Department of Microbiology, Mie University School of Medicine, Japan.
Med Microbiol Immunol. 1996 Sep;185(2):89-94. doi: 10.1007/s004300050019.
Interaction of the nucleocapsid (NP) and V proteins of human parainfluenza type 2 virus (HPIV-2) was investigated using a transient expression system. When the NP proteins were co-expressed with the V proteins, some of the NP proteins were translocated into the nuclei. These findings suggest that the NP protein interact with the V proteins. We examined the interaction of the NP proteins and the P, V proteins or deletion mutants of V protein using immunofluorescence and co-immunoprecipitation plus Western blotting analyses, and showed that the V proteins of HPIV-2 bind to the NP proteins and that the N-terminal domain of V protein interacts directly with the NP proteins. When the NP proteins were co-expressed with the V proteins or the N-terminal fragments (aa 1-46), the NP proteins were detected diffusely in the nuclei of the transfected cells, and were also detected in cytoplasmic inclusions. The NP and V proteins were co-localized in the nuclei or cytoplasm. Furthermore, the NP proteins were co-precipitated with the P, V, and V (1-164) proteins by a specific antibody. The P proteins interact more closely with the NP proteins than do the V proteins. These findings indicate that the V proteins have the ability to bind the NP proteins.
利用瞬时表达系统研究了人副流感病毒2型(HPIV-2)核衣壳(NP)蛋白与V蛋白之间的相互作用。当NP蛋白与V蛋白共表达时,部分NP蛋白会转运至细胞核内。这些发现表明NP蛋白与V蛋白存在相互作用。我们通过免疫荧光、免疫共沉淀及蛋白质免疫印迹分析,研究了NP蛋白与P蛋白、V蛋白或V蛋白缺失突变体之间的相互作用,结果显示HPIV-2的V蛋白可与NP蛋白结合,且V蛋白的N端结构域可直接与NP蛋白相互作用。当NP蛋白与V蛋白或V蛋白N端片段(第1至46位氨基酸)共表达时,在转染细胞的细胞核中可弥散性检测到NP蛋白,在细胞质内含物中也能检测到。NP蛋白和V蛋白在细胞核或细胞质中共定位。此外,特定抗体可使NP蛋白与P蛋白、V蛋白及V(1-164)蛋白发生共沉淀。P蛋白与NP蛋白的相互作用比V蛋白与NP蛋白的相互作用更为紧密。这些发现表明V蛋白具有结合NP蛋白的能力。