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Mokola virus (MOKV) is a nonsegmented, negative-sense RNA virus that belongs to the Lyssavirus genus and Rhabdoviridae family. MOKV phosphoprotein P is an essential component of the replication and transcription complex and acts as a cofactor for the viral RNA-dependent RNA polymerase. P recruits the viral polymerase to the nucleoprotein-bound viral RNA (N-RNA) via an interaction between its C-terminal domain and the N-RNA complex. Here we present a structure for this domain of MOKV P, obtained by expression of full-length P in Escherichia coli, which was subsequently truncated during crystallization. The structure has a high degree of homology with P of rabies virus, another member of Lyssavirus genus, and to a lesser degree with P of vesicular stomatitis virus (VSV), a member of the related Vesiculovirus genus. In addition, analysis of the crystal packing of this domain reveals a potential binding site for the nucleoprotein N. Using both site-directed mutagenesis and yeast two-hybrid experiments to measure P-N interaction, we have determined the relative roles of key amino acids involved in this interaction to map the region of P that binds N. This analysis also reveals a structural relationship between the N-RNA binding domain of the P proteins of the Rhabdoviridae and the Paramyxoviridae.

Original publication

DOI

10.1128/JVI.01520-09

Type

Journal article

Journal

J Virol

Publication Date

01/2010

Volume

84

Pages

1089 - 1096

Keywords

Binding Sites, Crystallization, Crystallography, X-Ray, Escherichia coli, Lyssavirus, Models, Molecular, Molecular Sequence Data, Mutagenesis, Site-Directed, Nucleocapsid Proteins, Phosphoproteins, Structure-Activity Relationship, Two-Hybrid System Techniques, Viral Proteins