Deciphering interactions used by the influenza virus NS1 protein to silence the host antiviral sensor protein RIG-I using a bacterial reverse two-hybrid system.
The majority of biological processes are controlled and regulated by an intricate network of thousands of interacting proteins. Identifying and understanding the key components of these protein networks, especially those that play a critical role in disease, is a challenge that promises to dramatically alter our current approach to healthcare. To facilitate this process, we have developed a method for the rapid construction of a chromosomally integrated, bacterial reverse two-hybrid system (RTHS) that enables the identification of interacting protein partners. Chromosomal integration of the RTHS enables stable protein expression, free of plasmid copy-number effects, as well as eliminating false positives arising from plasmid ejection. We have utilized this approach to identify the interactions used by the influenza virus NS1 protein to silence the host's antiviral defences.
School of Chemistry, University of Southampton, Southampton SO17 1BJ, UK.
Elena
E
Miranda
Fedor
F
Forafonov
Ali
A
Tavassoli
D000071457:DEAD Box Protein 58; D053487:DEAD-box RNA Helicases / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation; D023061:Gene Order; D054884:Host-Pathogen Interactions / Q000502:physiology*; D006801:Humans; D010957:Plasmids / Q000235:genetics; D011485:Protein Binding; D011971:Receptors, Immunologic; D020798:Two-Hybrid System Techniques; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism*
C085820:INS1 protein, influenza virus; D011971:Receptors, Immunologic; D017361:Viral Nonstructural Proteins; C452820:RIGI protein, human; D000071457:DEAD Box Protein 58; D053487:DEAD-box RNA Helicases