Induction of protein-protein interactions in live cells using light.
Protein-protein interactions are essential for many cellular processes. We have developed a technology called light-activated dimerization (LAD) to artificially induce protein hetero- and homodimerization in live cells using light. Using the FKF1 and GIGANTEA (GI) proteins of Arabidopsis thaliana, we have generated protein tags whose interaction is controlled by blue light. We demonstrated the utility of this system with LAD constructs that can recruit the small G-protein Rac1 to the plasma membrane and induce the local formation of lamellipodia in response to focal illumination. We also generated a light-activated transcription factor by fusing domains of GI and FKF1 to the DNA binding domain of Gal4 and the transactivation domain of VP16, respectively, showing that this technology is easily adapted to other systems. These studies set the stage for the development of light-regulated signaling molecules for controlling receptor activation, synapse formation and other signaling events in organisms.
Yazawa
Masayuki
M
Department of Neurobiology, Stanford University School of Medicine, Stanford, California, USA.
Sadaghiani
Amir M
AM
Hsueh
Brian
B
Dolmetsch
Ricardo E
RE
D000818:Animals; D029681:Arabidopsis Proteins / Q000378:metabolism; D002460:Cell Line; D006801:Humans; D007150:Immunohistochemistry; D008027:Light; D051379:Mice; D041681:NIH 3T3 Cells; D009479:Neuropeptides / Q000378:metabolism; D055668:Photochemical Processes; D055503:Protein Multimerization / Q000528:radiation effects; D011506:Proteins / Q000378:metabolism*; D011554:Pseudopodia / Q000378:metabolism; D014158:Transcription, Genetic / Q000528:radiation effects; D020744:rac GTP-Binding Proteins / Q000378:metabolism; D020830:rac1 GTP-Binding Protein
D029681:Arabidopsis Proteins; C410504:FKF1 protein, Arabidopsis; C121183:GI protein, Arabidopsis; D009479:Neuropeptides; D011506:Proteins; C484602:Rac1 protein, mouse; D020744:rac GTP-Binding Proteins; D020830:rac1 GTP-Binding Protein