Proximity-dependent biotinylation mediated by TurboID to identify protein-protein interaction networks in yeast.
J Cell Sci
; 132(11)2019 05 31.
Article
en En
| MEDLINE
| ID: mdl-31064814
ABSTRACT
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.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Saccharomyces cerevisiae
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Schizosaccharomyces
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Biotinilación
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Mapeo de Interacción de Proteínas
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Mapas de Interacción de Proteínas
Idioma:
En
Revista:
J Cell Sci
Año:
2019
Tipo del documento:
Article