Poster abstracts

Poster number 43 submitted by Francis Fordjour

Characterization of the N-terminal Domain of Ebolavirus Nucleoprotein

Francis Kusi Fordjour (Chemistry & Biochemistry, DePauw University), Lauren Salay (Chemistry & Biochemistry, DePauw University), Jonathan M. Howard (Chemistry & Biochemistry, DePauw University), Lauren M. Guggina (Chemistry & Biochemistry, DePauw University), W. Maxwell Hudson (Chemistry & Biochemistry, DePauw University), Sharon Crary (Chemistry & Biochemistry, DePauw University)

Abstract:
Ebola virus is a negative sense RNA virus that can cause viral hemorrhagic fever. The nucleocapsid structure includes the RNA and several viral proteins, with the nucleoprotein (NP) directly interacting with the RNA. To characterize the mechanism involved in this RNA-protein interaction we are establishing a source of pure, soluble NP. Using protein secondary structure prediction software (Poodle, PredictProtein, and PSIPRED) we identified the C-terminus of NP as likely to be intrinsically disordered and thus a potential culprit for the aggregation seen during previous NP purification attempts. We therefore constructed plasmids which coded for the N-terminus of NP with various C-terminal truncations for bacterial overexpression. Truncating NP at amino acid 356 resulted in purification of soluble protein, even after cleavage of the maltose binding protein used to stabilize NP during the expression and purification process. Experiments to study the interactions of NP with the viral RNA, as well as efforts to determine the structure of NP, are being pursued.

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Keywords: Ebolavirus, Nucleoprotein, RNA