BOTH THE N-TERMINAL AND THE C-TERMINAL DOMAINS OF THE NOMINAL PHOSPHOPROTEIN OF RABIES VIRUS ARE INVOLVED IN BINDING TO THE NUCLEOPROTEIN
BOTH THE N-TERMINAL AND THE C-TERMINAL DOMAINS OF THE NOMINAL PHOSPHOPROTEIN OF RABIES VIRUS ARE INVOLVED IN BINDING TO THE NUCLEOPROTEIN
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DOI：
10.1006/viro.1994.1222
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发表时间：
1994-05-01
期刊：
影响因子：
3.7
通讯作者：
DIETZSCHOLD, B
中科院分区：
文献类型：
--
作者：
FU, ZF;ZHENG, YM;DIETZSCHOLD, B
通过在昆虫细胞和体外转录-翻译偶联系统中表达狂犬病病毒（Evelyn Rokitnicki Abelseth株）的名义磷蛋白（NS）和核蛋白（N），研究了它们之间的相互作用。在昆虫细胞中单独表达的N和NS蛋白在体外相互作用，并以与狂犬病病毒体中发现的相似的N：NS比例形成复合物。在体外转录-翻译系统中，N和NS蛋白在同时合成时形成N-NS复合物，该复合物可以与抗N或抗NS抗体免疫沉淀。NS突变蛋白的C-末端缺失多达166个氨基酸，仍然能够形成复合物时，同时合成的N蛋白。然而，当单独合成NS突变蛋白和N蛋白然后混合在一起时，只有完整的NS蛋白和从C-末端缺失24个氨基酸的NS突变蛋白与N蛋白结合，而从NS蛋白的C-末端缺失47个或更多个氨基酸导致与N蛋白的结合完全丧失。当同时或单独合成时，N-末端缺失多达68个氨基酸的NS突变体与N蛋白结合。这些结果表明，狂犬病病毒NS蛋白的N-和C-末端结构域参与狂犬病病毒N蛋白的结合，但不是以相互依赖的方式。NS蛋白的N-末端结构域与N蛋白的相互作用仅在两种蛋白同时合成期间发生，而当这两种蛋白同时或单独合成时，NS蛋白的C-末端区域可以与N蛋白结合。NS蛋白与N蛋白的两个结合位点可能在调节病毒转录和复制以及病毒组装中具有重要功能，（C）1994 Academic Press，Inc.
The interaction of the nominal phosphoprotein (NS) and the nucleoprotein (N) of rabies virus (Evelyn Rokitnicki Abelseth strain) was investigated by expressing these proteins in insect cells and in an in vitro coupled transcription-translation system. The N and NS proteins individually expressed in insect cells interacted with each other in vitro and formed complexes at a N:NS ratio similar to those found in rabies virions. In the in vitro transcription-translation system, both the N and NS proteins when systhesized simultaneously formed N-NS complexes which could be immunoprecipitated with either anti-N or anti-NS antibodies. NS mutant proteins with C-terminal deletion of up to 166 amino acids were still able to form complexes with N protein when synthesized simultaneously. However, when the NS mutant proteins and the N protein were synthesized individually and then mixed together, only the intact NS protein and NS mutant protein with 24 amino acids deleted from the C-terminus bound to the N protein, whereas deletion of 47 or more amino acids from the C-terminus of the NS protein resulted in total loss of binding to the N protein. NS mutants with N-terminal deletions of up to 68 amino acids bound to the N protein when synthesized either simultaneously or individually. These results indicate that both the N- and C-terminal domains of the NS protein of rabies virus are involved in the binding to rabies virus N protein but not in a mutually dependent manner. The interaction of the N-terminal domain of the NS protein with N protein occurs only during simultaneous synthesis of both proteins, whereas the C-terminal region of the NS protein can bind to the N protein when these two proteins are synthesized either simultaneously or separately. The two binding sites of the NS protein to N protein might have important functions in regulating virus transcription and replication as well as in virus assembly, (C) 1994 Academic Press, Inc.