Conformational Changes in the Connector Protein Complex of the Bacteriophage φ29 DNA Packaging Motor.pdfVIP

Computational and Mathematical Methods in Medicine Vol. 9, Nos. 3–4, September–December 2008, 327–337 Conformational changes in the connector protein complex of the bacteriophage f29 DNA packaging motor Arron C. Tolley and Nicola J. Stonehouse* Astbury Centre for Structural Molecular Biology and Faculty of Biological Sciences, University of Leeds, Leeds, UK ( Received 4 February 2008; final version received 29 March 2008 ) DNA packaging in the bacteriophage f29 involves a molecular motor. It is proposed that dsDNA is packaged through a channel in a connector located at the 5-fold vertex of a preformed prolate icosahedral capsid. The packaging motor also consists of virally-encoded RNA molecules (pRNA) coupled to ATPases. Data obtained from studies using surface plasmon resonance, fluorescence quenching and circular dichroism are presented to demonstrate the importance of the N-termini of the connector protein subunits in pRNA interaction and in conformational change. Based on our findings, we propose a model of DNA packaging based on connector conformational change. Keywords: RNA–protein interaction; bacteriophage f29; molecular motor; virus assembly; conformational change 1. Introduction Viruses have been identified in a wide range of organisms and all utilize a protein shell or capsid for the protection of their nucleic acid genome. The genome packaging event is therefore a key step in the lifecycle of every virus. In order to understand how these processes occur, it is important to dissect the fine details of the assembly pathway and characterize intermediates formed during nucleic acid packaging. Furthermore, an understandi