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Mapping the proximity interaction network of the RHO-family GTPases reveals signalling pathways and regulatory mechanisms
First Author: Halil Bagci
  TGuanine nucleotide exchange factors (RHOGEFs) and GTPase activating proteins (RHOGAPs) coordinate the activation state of RHO-family GTPases for binding to effectors. Here, we exploit proximity-dependent biotinylation (BioID) to systematically define the RHO-family proximity interaction network from 28 baits producing 9939 high-confidence proximity interactions in two cell lines. Exploiting the nucleotide states of RHO GTPases, we revealed the landscape of interactions with RHOGEFs/RHOGAPs. We systematically defined effectors of RHO proteins revealing candidates for classical and atypical RHOs. We used optogenetics to demonstrate that KIAA0355/GARRE is a RAC1 interactor. A functional screen of RHOG candidate effectors identified PLEKHG3 as a promoter of RAC-mediated membrane ruffling downstream of RHOG. We identified that active RHOA binds the kinase SLK in Drosophila and mammalian cells to promote EZRIN/RADIXIN/MOESIN phosphorylation. Our proximity interactions data paves the way for dissecting additional RHO signalling pathways and the approaches described here are applicable to the RAS-family.

Please contact Jean-François Côté ( for details.

Navigating this website

• The data included here is by default filtered with AvgP ≥ 0.95, as calculated by SAINTexpress; unselect the "high confidence" buttons to view all data. You can also download a complete interactome table through the Supplementary data section (left).

Select "Explore baits" (left) to view all baits profiled, organized alphabetically (view as a list). Clicking on a bait will retrieve all its interactors (high confidence by default) and indicate which interactions are already deposited in the BioGRID and IntAct databases.

• Selecting multiple baits (or all baits) will enable a download of the SAINT result file that can be opened at ProHits-viz for visualization and further exploration.

• Also see the many links to excellent external resources for the baits and/or preys, including ProteinAtlas, ProteomicsDB, NCBI Gene, GeneCards and UniProt.

• Search for a bait or prey by Official Gene Symbol through the "Search" tab on left (or at the top). You will be able to search across all projects you have access to.

• The manuscript is in press in Nature Cell Biology. The submitted version of the manuscript will be associated with Supplementary Data containing additional figures, tables and input files for Cytoscape and ProHits-viz (not yet enabled).

Note that the mass spectrometry data has been deposited at MassIVE.