Abstract
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 non-functional 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.
Original language | English |
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Pages (from-to) | 108-111 |
Number of pages | 4 |
Journal | Journal of Virological Methods |
Volume | 176 |
Issue number | 1-2 |
DOIs | |
State | Published - Sep 2011 |
Bibliographical note
Funding Information:We thank Elizabeth Kolb for editing the manuscript and thank members of the Li lab for helpful discussions and critical reviews of the manuscript. We thank Ruben Donis at the CDC for providing Flu B reagents. We acknowledge the use of the SDSU-Functional Genomics Core Facility supported in part by NSF/EPSCoR Grant No. 0091948, the Center of Excellence in Drought Tolerance through the South Dakota 2010 Initiative and the South Dakota Agri. Exp. Station. Research in the S.C. laboratory is supported by the South Dakota Agri. Exp. Station and the South Dakota 2010 Research Center, BCAAP (Biological Control and Analysis of Applied Photonics) Fund (3SG163 to S.C.). This work was supported by the South Dakota Agricultural Experiment Station (3AH203 to F.L.) and Public Health Service grants (AI078177) from NIAID to F.L.
Keywords
- Protein-protein interaction
- Split luciferase complementation assay
- Viral disease
ASJC Scopus subject areas
- Virology