Substitution of two residues in the measles virus nucleoprotein results in an impaired self-association

被引:70
作者
Karlin, D
Longhi, S
Canard, B
机构
[1] Univ Aix Marseille 1, CNRS, AFMB, ESIL, F-13288 Marseille 09, France
[2] Univ Aix Marseille 1, ESIL, F-13288 Marseille, France
[3] UMR 6098 CNRS, F-13288 Marseille 09, France
关键词
Paramyxovindae; morbillivirus; measles virus; nucleocapsid; nucleoproteins; point mutation; phosphoproteins; protein-protein interactions; binding sites; electron microscopy;
D O I
10.1006/viro.2002.1634
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The nucleoprotein (N) of measles virus encapsidates viral genomic RNA to form a helical nucleocapsid. Its strong self-association is a major hurdle in determining its high-resolution structure using X-ray crystallography We report the bacterial expression, purification, and characterization of a variant N that has lost its ability to form nucleocapsid-like structures after substitution of two residues by polar residues. Using immunoprecipitation, circular dichroism, and limited proteolysis studies, we show that this nucleoprotein retains a folding similar to wild-type N. Furthermore, the variant N binds the phosphoprotein, indicating that it retains biochemical relevance. We also present evidence indicating that the N-terminus of N lies at the surface of the nucleocapsid. Beyond the identification of one region of N involved in self-association, our results should facilitate structural studies of N using X-ray crystallography. (C) 2002 Elsevier Science (USA).
引用
收藏
页码:420 / 432
页数:13
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