Barcode fusion genetics-protein-fragment complementation assay (BFG-PCA): tools and resources that expand the potential for binary protein interaction discovery. Issue 9 (7th February 2022)

• Record Type: Journal Article
• Authors: Evans-Yamamoto, Daniel; Rouleau, François D; Nanda, Piyush; Makanae, Koji; Liu, Yin; Després, Philippe C; Matsuo, Hitoshi; Seki, Motoaki; Dubé, Alexandre K; Ascencio, Diana; Yachie, Nozomu; Landry, Christian R
• Is Part Of: Nucleic acids research. Volume 50:Issue 9(2022)
• Page Start: e54
• Page End: e54
• Publication Date: 2022-02-07
• Subjects: Nucleic acids -- Periodicals; Molecular biology -- Periodicals; 572.805
• DOI: 10.1093/nar/gkac045
• Languages: English
• ISSNs: 0305-1048

Abstract

Abstract: Barcode fusion genetics (BFG) utilizes deep sequencing to improve the throughput of protein–protein interaction (PPI) screening in pools. BFG has been implemented in Yeast two-hybrid (Y2H) screens (BFG-Y2H). While Y2H requires test protein pairs to localize in the nucleus for reporter reconstruction, dihydrofolate reductase protein-fragment complementation assay (DHFR-PCA) allows proteins to localize in broader subcellular contexts and proves to be largely orthogonal to Y2H. Here, we implemented BFG to DHFR-PCA (BFG-PCA). This plasmid-based system can leverage ORF collections across model organisms to perform comparative analysis, unlike the original DHFR-PCA that requires yeast genomic integration. The scalability and quality of BFG-PCA were demonstrated by screening human and yeast interactions for >11 000 bait-prey pairs. BFG-PCA showed high-sensitivity and high-specificity for capturing known interactions for both species. BFG-Y2H and BFG-PCA capture distinct sets of PPIs, which can partially be explained based on the domain orientation of the reporter tags. BFG-PCA is a high-throughput protein interaction technology to interrogate binary PPIs that exploits clone collections from any species of interest, expanding the scope of PPI assays.