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Article Dans Une Revue Cells Année : 2023

A Live Cell Protein Complementation Assay for ORFeome-Wide Probing of Human HOX Interactomes

Résumé

Biological pathways rely on the formation of intricate protein interaction networks called interactomes. Getting a comprehensive map of interactomes implies the development of tools that allow one to capture transient and low-affinity protein–protein interactions (PPIs) in live conditions. Here we presented an experimental strategy: the Cell-PCA (cell-based protein complementation assay), which was based on bimolecular fluorescence complementation (BiFC) for ORFeome-wide screening of proteins that interact with different bait proteins in the same live cell context, by combining high-throughput sequencing method. The specificity and sensitivity of the Cell-PCA was established by using a wild-type and a single-amino-acid-mutated HOXA9 protein, and the approach was subsequently applied to seven additional human HOX proteins. These proof-of-concept experiments revealed novel molecular properties of HOX interactomes and led to the identification of a novel cofactor of HOXB13 that promoted its proliferative activity in a cancer cell context. Taken together, our work demonstrated that the Cell-PCA was pertinent for revealing and, importantly, comparing the interactomes of different or highly related bait proteins in the same cell context.
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Dates et versions

hal-04176609 , version 1 (03-08-2023)

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Yunlong Jia, Jonathan Reboulet, Benjamin Gillet, Sandrine Hughes, Christelle Forcet, et al.. A Live Cell Protein Complementation Assay for ORFeome-Wide Probing of Human HOX Interactomes. Cells, 2023, 12 (1), pp.200. ⟨10.3390/cells12010200⟩. ⟨hal-04176609⟩
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