Proximity-dependent biotinylation mediated by TurboID to identify protein-protein interaction networks in yeast.
The use of proximity-dependent biotinylation assays coupled to mass spectrometry (PDB-MS) has changed the field of protein-protein interaction studies. However, despite the recurrent and successful use of BioID-based protein-protein interactions screening in mammalian cells, the implementation of PDB-MS in yeast has not been effective. Here, we report a simple and ... rapid approach in yeast to effectively screen for proximal and interacting proteins in their natural cellular environment by using TurboID, a recently described version of the BirA biotin ligase. Using the protein arginine methyltransferase Rmt3 and the RNA exosome subunits, Rrp6 and Dis3, the application of PDB-MS in yeast by using TurboID was able to recover protein-protein interactions previously identified using other biochemical approaches and provided new complementary information for a given protein bait. The development of a rapid and effective PDB assay that can systematically analyze protein-protein interactions in living yeast cells opens the way for large-scale proteomics studies in this powerful model organism.
Mesh Terms:
Biotinylation, Carbon-Nitrogen Ligases, Exosome Multienzyme Ribonuclease Complex, Mass Spectrometry, Protein Interaction Mapping, Protein Interaction Maps, Protein-Arginine N-Methyltransferases, Proteomics, Ribonucleases, Saccharomyces cerevisiae, Schizosaccharomyces, Schizosaccharomyces pombe Proteins
Biotinylation, Carbon-Nitrogen Ligases, Exosome Multienzyme Ribonuclease Complex, Mass Spectrometry, Protein Interaction Mapping, Protein Interaction Maps, Protein-Arginine N-Methyltransferases, Proteomics, Ribonucleases, Saccharomyces cerevisiae, Schizosaccharomyces, Schizosaccharomyces pombe Proteins
J Cell Sci
Date: Dec. 31, 2018
PubMed ID: 31064814
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