Isolation of RBP47 interactome

Stress granules are dynamic structures, in which components belonging to the shell structure can exchange rapidly while components in the core layer may be less dynamic. The last group of proteins has been suggested to act as a scaffold to which other SG components are recruited. In a previous study, we observed that TSN does not exchange between the cytoplasm and foci upon a fluorescence recovery after photobleaching (FRAP) analysis, suggesting a role as a scaffold protein. Deletion of scaffold-like molecules has been suggest to have a high impact in membraneles organelles composition. Accordingly, and to better understand the role of TSN as a platform for SG components in plants, we characterized the SG proteome by isolation of GFP-tagged RBP47 protein from WT and double tsn1 tsn2 knock-out expressing plants.