Coronaviruses

Pierre-Olivier Vidalain, Yves Jacob, Marne C Hagemeijer, Louis M Jones, Grégory Neveu, Jean-Pierre Roussarie, Peter J M Rottier, Frédéric Tangy, Cornelis A M de Haan, Marne C. Hagemeijer, Louis M. Jones, Peter J. M. Rottier, Cornelis A. M. de Haan, Vidalain, Pierre-Olivier, Jacob, Yves, Hagemeijer, Marne C., Jones, Louis M., Neveu, Grégory, Roussarie, Jean-Pierre, Rottier, Peter J. M., Tangy, Frédéric, de Haan, Cornelis A. M., Haan, Cornelis A. M.

Helena Jane Maier, Erica Bickerton, Paul Britton

Over the last 2 decades, yeast two-hybrid became an invaluable technique to decipher protein-protein interaction networks. In the field of virology, it has proven instrumental to identify virus-host interactions that are involved in viral embezzlement of cellular functions and inhibition of immune mechanisms. Here, we present a yeast two-hybrid protocol that has been used in our laboratory since 2006 to search for cellular partners of more than 300 viral proteins. Our aim was to develop a robust and straightforward pipeline, which minimizes false-positive interactions with a decent coverage of target cDNA libraries, and only requires a minimum of equipment. We also discuss reasons that motivated our technical choices and compromises that had to be made. This protocol has been used to screen most non-structural proteins of murine hepatitis virus (MHV), a member of betacoronavirus genus, against a mouse brain cDNA library. Typical results were obtained and are presented in this report.