Structure of the nucleoprotein binding domain of Mokola virus phosphoprotein.

Journal article

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.

Authors: Assenberg, R; Delmas, O; Ren, J; Vidalain, P; Verma, A

• Publication status: Published
• Journal: Journal of virology
• Volume: 84
• Issue: 2
• Pages: 1089-1096
• Publication date: 2010-01-01
• DOI: 10.1128/jvi.01520-09
• EISSN: 1098-5514
• ISSN: 0022-538X
• Language: English
• Keywords: Two-Hybrid System Techniques; Crystallography, X-Ray; Molecular Sequence Data; Models, Molecular; Nucleocapsid Proteins; Structure-Activity Relationship; Escherichia coli; Phosphoproteins; Lyssavirus; Binding Sites; Mutagenesis, Site-Directed; Crystallization; Viral Proteins