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

Yunlong Jia, Jonathan Reboulet, Benjamin Gillet, View ORCID ProfileSandrine Hughes, View ORCID ProfileChristelle Forcet, Violaine Tribollet, View ORCID ProfileNawal Hadj Sleiman, Cindy Kundlacz, Jean-Marc Vanacker, Françoise Bleicher, View ORCID ProfileSamir Merabet
doi: https://doi.org/10.1101/2022.11.09.515750
Yunlong Jia
1IGFL, CNRS UMR5242, ENS-Lyon, UCBL-1, INRA USC1370, 32 Av. Tony Garnier, 69007 Lyon, France
2Department of Developmental and Cell Biology, University of California, Irvine, Irvine, CA 92697, USA
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Jonathan Reboulet
1IGFL, CNRS UMR5242, ENS-Lyon, UCBL-1, INRA USC1370, 32 Av. Tony Garnier, 69007 Lyon, France
3LiPiCs, 46 Allée d’Italie, 69007 Lyon, France
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Benjamin Gillet
1IGFL, CNRS UMR5242, ENS-Lyon, UCBL-1, INRA USC1370, 32 Av. Tony Garnier, 69007 Lyon, France
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Sandrine Hughes
1IGFL, CNRS UMR5242, ENS-Lyon, UCBL-1, INRA USC1370, 32 Av. Tony Garnier, 69007 Lyon, France
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  • ORCID record for Sandrine Hughes
Christelle Forcet
1IGFL, CNRS UMR5242, ENS-Lyon, UCBL-1, INRA USC1370, 32 Av. Tony Garnier, 69007 Lyon, France
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  • ORCID record for Christelle Forcet
Violaine Tribollet
1IGFL, CNRS UMR5242, ENS-Lyon, UCBL-1, INRA USC1370, 32 Av. Tony Garnier, 69007 Lyon, France
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Nawal Hadj Sleiman
1IGFL, CNRS UMR5242, ENS-Lyon, UCBL-1, INRA USC1370, 32 Av. Tony Garnier, 69007 Lyon, France
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  • ORCID record for Nawal Hadj Sleiman
Cindy Kundlacz
1IGFL, CNRS UMR5242, ENS-Lyon, UCBL-1, INRA USC1370, 32 Av. Tony Garnier, 69007 Lyon, France
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Jean-Marc Vanacker
1IGFL, CNRS UMR5242, ENS-Lyon, UCBL-1, INRA USC1370, 32 Av. Tony Garnier, 69007 Lyon, France
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Françoise Bleicher
1IGFL, CNRS UMR5242, ENS-Lyon, UCBL-1, INRA USC1370, 32 Av. Tony Garnier, 69007 Lyon, France
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  • For correspondence: samir.merabet@ens-lyon.fr françoise.bleicher@univ-lyon1.fr
Samir Merabet
1IGFL, CNRS UMR5242, ENS-Lyon, UCBL-1, INRA USC1370, 32 Av. Tony Garnier, 69007 Lyon, France
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  • ORCID record for Samir Merabet
  • For correspondence: samir.merabet@ens-lyon.fr françoise.bleicher@univ-lyon1.fr
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Abstract

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 proof-of-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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Posted November 09, 2022.
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A live cell protein complementation assay for ORFeome-wide probing of human HOX interactomes
Yunlong Jia, Jonathan Reboulet, Benjamin Gillet, Sandrine Hughes, Christelle Forcet, Violaine Tribollet, Nawal Hadj Sleiman, Cindy Kundlacz, Jean-Marc Vanacker, Françoise Bleicher, Samir Merabet
bioRxiv 2022.11.09.515750; doi: https://doi.org/10.1101/2022.11.09.515750
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A live cell protein complementation assay for ORFeome-wide probing of human HOX interactomes
Yunlong Jia, Jonathan Reboulet, Benjamin Gillet, Sandrine Hughes, Christelle Forcet, Violaine Tribollet, Nawal Hadj Sleiman, Cindy Kundlacz, Jean-Marc Vanacker, Françoise Bleicher, Samir Merabet
bioRxiv 2022.11.09.515750; doi: https://doi.org/10.1101/2022.11.09.515750

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