Interaction between nucleocapsid protein (NP) and phosphoprotein (P) of human parainfluenza virus type 2: one of the two NP binding sites on P is essential for granule formation.
Interaction between nucleocapsid protein (NP) and phosphoprotein (P) of human parainfluenza virus type 2: one of the two NP binding sites on P is essential for granule formation.
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DOI：
10.1099/0022-1317-77-10-2457
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发表时间：
1996-10
期刊：
影响因子：
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通讯作者：
M. Nishio;M. Tsurudome;Mitsuo Kawano;Noriko Watanabe;S. Ohgimoto;Morihiro Ito;H. Komada;Yasuhiko Ito
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文献类型：
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作者：
M. Nishio;M. Tsurudome;Mitsuo Kawano;Noriko Watanabe;S. Ohgimoto;Morihiro Ito;H. Komada;Yasuhiko Ito
副粘病毒磷酸 (P) 和核衣壳 (NP) 蛋白参与病毒基因组的转录和复制。为了研究 NP 和 P 蛋白之间的相互作用，我们建立了组成型表达人副流感病毒 2 型 (hPIV-2) NP 和/或 P 蛋白的 HeLa 细胞系。免疫共沉淀分析表明，NP 和 P 蛋白可以在表达这两种蛋白的 HeLa 细胞（HeLa-NP+P 细胞）以及 HeLa-NP 和 HeLa-P 细胞的混合细胞裂解物中形成复合物。对 P 蛋白进行缺失突变体分析，以确定 P 蛋白与 NP 蛋白相互作用的区域。结果表明，P蛋白上存在两个独立的NP结合位点：一个位于该蛋白的N端部分，aa 1-47，另一个位于该蛋白的C端部分，aa 357-395。此外，共表达具有 N 末端缺失的 NP 和 P 蛋白的细胞显示出与 hPIV-2 感染细胞中发现的类似的免疫荧光染色模式（颗粒模式）。然而，共表达具有 C 末端缺失的 NP 和 P 蛋白的细胞显示出不同的免疫荧光染色模式（弥散模式），表明 C 末端区域是颗粒形成所必需的。
The paramyxovirus phospho- (P) and nucleocapsid (NP) proteins are involved in transcription and replication of the viral genome. To study the interaction between NP and P proteins, we established HeLa cell lines that constitutively expressed the NP and/or P proteins of human parainfluenza virus type 2 (hPIV-2). Co-immunoprecipitation assays revealed that the NP and P proteins can form complexes in HeLa cells expressing both proteins (HeLa-NP+P cells) and in mixed cell lysates of HeLa-NP and HeLa-P cells. Deletion mutant analysis of the P protein was performed to identify the regions of P protein that interact with NP protein. The results indicate that two independent NP-binding sites exist on P protein: one is located in the N-terminal part of the protein, aa 1-47, and the other in the C-terminal part, aa 357-395. In addition, cells co-expressing NP and P proteins with N-terminal deletions showed immunofluorescence staining patterns (granular pattern) similar to those found in hPIV-2-infected cells. However, cells co-expressing NP and P proteins with C-terminal deletions showed a different immunofluorescence staining pattern (diffuse pattern), indicating that the C-terminal region is required for granule formation.