Biology subjects

Reboulet, J.

Publications and source records attributed to Reboulet, J..

3 recordsLinked to original sources

bioRxiv2023

Optimization for High-Throughput BiFC screening

The Cell-PCA screen, since its inception, has provided an efficient method for analyzing cellular interactomes and has been used in various biological studies involving proteins like MYC, PER2, and ERK. With rapid advancements in biotechnology, including tools for protein function investigation, the Cell-PCA screen remains relevant. However, despite its successful application in recent studies, there are areas for optimization to ensure its continued relevance in the face of evolving technological advancements.

bioengineering↗

bioRxiv2022

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

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.

molecular biology↗

bioRxiv2019

Role of a versatile peptide motif in controlling Hox nuclear export and autophagy in the Drosophila fat body

Hox proteins are major regulators of embryonic development, acting in the nucleus to regulate the expression of their numerous downstream target genes. By analyzing deleted forms of the Drosophila Hox protein Ultrabithorax (Ubx), we revealed the presence of an unconventional Nuclear Export Signal (NES) that overlaps with the highly conserved hexapeptide (HX) motif. This short linear motif was originally described as mediating the interaction with the PBC proteins, a generic and crucial class of Hox transcriptional cofactors in development and cancer. Here we show that the HX motif is involved in the interaction with the major CRM1/Embargoed exportin protein. This novel role was found in several Drosophila and human Hox proteins. We provide evidence that HX-dependent Hox nuclear export is tightly regulated in the Drosophila fat body to control the onset of autophagy. Our results underline the high molecular versatility of a unique short peptide motif for controlling context-dependent activity of Hox proteins at both transcriptional and non-transcriptional levels.

developmental biology↗