Genome-wide physical protein±protein interaction(PPI)mapping remains a major challenge for current technologies.Here,we reported a high-efficiency BiFC-seq method,yeastenhanced green fluorescent protein-based bim...Genome-wide physical protein±protein interaction(PPI)mapping remains a major challenge for current technologies.Here,we reported a high-efficiency BiFC-seq method,yeastenhanced green fluorescent protein-based bimolecular fluorescence complementation(y EGFPBiFC)coupled with next-generation DNA sequencing,for interactome mapping.We first applied y EGFP-BiFC method to systematically investigate an intraviral network of the Ebola virus.Two-thirds(9/14)of known interactions of EBOV were recaptured,and five novel interactions were discovered.Next,we used the BiFC-seq method to map the interactome of the tumor protein p53.We identified 97 interactors of p53,more than three-quarters of which were novel.Furthermore,in a more complex background,we screened potential interactors by pooling two BiFC libraries together and revealed a network of 229 interactions among 205 proteins.These results show that BiFC-seq is a highly sensitive,rapid,and economical method for genome-wide interactome mapping.展开更多
基金supported by grants from the National Key R&D Program of China(Grant No.2017YFA0505700)the National Key Lab of Proteomics of China(Grant Nos.SKLP-K201805,SKLP-K201804,and SKLP-Y201703)。
文摘Genome-wide physical protein±protein interaction(PPI)mapping remains a major challenge for current technologies.Here,we reported a high-efficiency BiFC-seq method,yeastenhanced green fluorescent protein-based bimolecular fluorescence complementation(y EGFPBiFC)coupled with next-generation DNA sequencing,for interactome mapping.We first applied y EGFP-BiFC method to systematically investigate an intraviral network of the Ebola virus.Two-thirds(9/14)of known interactions of EBOV were recaptured,and five novel interactions were discovered.Next,we used the BiFC-seq method to map the interactome of the tumor protein p53.We identified 97 interactors of p53,more than three-quarters of which were novel.Furthermore,in a more complex background,we screened potential interactors by pooling two BiFC libraries together and revealed a network of 229 interactions among 205 proteins.These results show that BiFC-seq is a highly sensitive,rapid,and economical method for genome-wide interactome mapping.
