Characterisation of the RNA binding properties of the coronavirus infectious bronchitis virus nucleocapsid protein amino-terminal region - Analytical Methods & Tools for Lyophilization

Characterisation of the RNA binding properties of the coronavirus infectious bronchitis virus nucleocapsid protein amino-terminal region
Received 20 July 2006; revised 5 September 2006; accepted 22 September 2006. Available online 2 October 2006.
Kelly-Anne Spencera and Julian A. Hiscoxa, b
Edited by Gianni Cesareni
FEB Letters
ScienceDirect
Copyright ? 2006 Published by Elsevier B.V.
aInstitute of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds LS2 9JT, UK
bAstbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK
Abstract
The coronavirus nucleocapsid (N) protein binds viral RNA to form the ribonucleocapsid and regulate RNA synthesis. The interaction of N protein with viral RNA was investigated using circular dichroism and surface plasmon resonance. N protein underwent a conformational change upon binding viral RNA and the data indicated electrostatic interactions were involved in the binding of the protein to RNA. Kinetic analysis suggested the amino-terminal region facilitates long-range non-specific interactions between N protein and viral RNA, thus bringing the RNA into close proximity to N protein allowing specific contacts to form via a ?lure? and ?lock? mechanism.
Keywords: Coronavirus; RNA binding; Nucleocapsid protein; IBV; Surface plasmon resonance; Circular dichroism
Corresponding author. Address: Institute of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds LS2 9JT, UK. Fax: +44 113 343 3167.
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