Properties of biomolecular condensates defined by the receptor independent activator of G protein Signaling 3 (AGS3).

Activator of G-protein Signaling 3 (AGS3), a receptor independent activator of G-protein signaling, oscillates among defined subcellular compartments and biomolecular condensates (BMCs) in a regulated manner that is likely related to the functional diversity of the protein. We determined the influence of cell stress on the cellular distribution of AGS3 and core material properties of AGS3 BMCs. Cellular stress (oxidative, pHi, thermal) induced the formation of AGS3 BMCs in Hela and COS-7 cells as determined by fluorescent microscopy. Oxidative stress-induced AGS3-BMCs were distinct from G3BP1 stress granules and from RNA processing BMCs defined by the P-body protein Dcp1a. Immunoblots indicated that cellular stress shifted AGS3, but not the stress granule protein G3BP1 to a membrane pellet fraction following cell lysis. The stress-induced generation of AGS3 BMCs was reduced by co-expression of the signaling protein Gαi3, but not the AGS3 binding partner DVL2. Fluorescent recovery following photobleaching of individual AGS3-BMCs indicated distinct diffusion kinetics and restricted fluidity for AGS3-BMCs. These data suggest that AGS3 BMCs represents a distinct class of stress granules that serve as a previously unrecognized signal processing node.