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Pré-Publication, Document De Travail Année : 2022

A live cell protein complementation assay for ORFeome-wide probing of human HOX interactomes

Yunlong Jia
  • Fonction : Auteur
Jonathan Reboulet
  • Fonction : Auteur
Benjamin Gillet
  • Fonction : Auteur
Sandrine Hughes
Christelle Forcet
Violaine Tribollet
  • Fonction : Auteur
Nawal Hadj Sleiman
Cindy Kundlacz
  • Fonction : Auteur
Jean-Marc Vanacker
  • Fonction : Auteur
Françoise Bleicher
  • Fonction : Auteur
Samir Merabet

Résumé

Biological pathways rely on the formation of intricate protein interaction networks called interactomes. Getting a comprehensive map of interactomes implies developing tools that allow capturing transient and low affinity protein-protein interactions (PPIs) in live conditions. Here we present an experimental strategy, Cell-PCA (Cell Protein Complementation Assay), which is based on BiFC (Bimolecular Fluorescence Complementation) and high throughput sequencing for ORFeome-wide analyses of different interactomes in the same live cell context. The specificity and sensitivity of Cell-PCA was established by using a wild type and a single amino-acid mutated HOXA9 protein, and the approach was subsequently applied for seven additional human HOX proteins. These proofof-concept experiments revealed novel molecular properties of HOX interactomes and led to the identification of a novel cofactor of HOXB13 for promoting its proliferative activity in a cancer cell context. Taken together, our work demonstrates that Cell-PCA is pertinent for revealing and, importantly, comparing interactomes between different or highly related bait proteins in the same cell context.
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Dates et versions

hal-03869735 , version 1 (24-11-2022)

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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. 2022. ⟨hal-03869735⟩
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