Intraviral protein-protein interactions are critical for virus survival in the host. Discovery of such interactions is important to understand molecular mechanisms of viral replication and pathogenesis. The development of a cell-based assay that can be employed to examine systematically viral protein interactions is described. The method, known as the Split Luciferase Complementation Assay (SLCA), is based on the principle that N- and C-terminal domains of luciferase alone do not emit luminescence; however, if fused to interacting proteins the two nonfunctional halves can be brought into close enough proximity through a specific protein-protein interaction to restore the functions of the enzyme and emit detectable light. The well-studied influenza B polymerase acidic protein (PA) and basic protein 1 (PB1) interaction was used as a model system to develop the assay. Consistent with previous studies, a strong PA-PB1 interaction was demonstrated in the assay. The PA-PB1 interaction was also disrupted by single amino acid mutations in the N-terminal domain of PB1 that is responsible for binding PA. The described SLCA is highly specific and easy to perform, and thus may be useful for studying protein-protein interactions in viral diseases.
Journal of Virological Methods
DOI of Published Version
Deng, Qiji; Wang, Dan; Xiang, Xiaoxiao; Hardwidge, Philip R.; Kaushik, Radhey S.; Wolff, Thorsten; Chakravarty, Suvobrata; and Li, Feng, "Application of a Split Luciferase Complementation Assay for theDetection of Viral Protein-Protein Interactions" (2011). Chemistry and Biochemistry Faculty Publications. 22.