title: Optogenetic engineering: light-directed cell motility.
abstract: Genetically encoded, light-activatable proteins provide the means to probe biochemical pathways at specific subcellular locations with exquisite temporal control. However, engineering these systems in order to provide a dramatic jump in localized activity, while retaining a low dark-state background remains a significant challenge. When placed within the framework of a genetically encodable, light-activatable heterodimerizer system, the actin-remodelling protein cofilin induces dramatic changes in the F-actin network and consequent cell motility upon illumination. We demonstrate that the use of a partially impaired mutant of cofilin is critical for maintaining low background activity in the dark. We also show that light-directed recruitment of the reduced activity cofilin mutants to the cytoskeleton is sufficient to induce F-actin remodeling, formation of filopodia, and directed cell motility.
authors.affiliation: Department of Chemistry, Division of Chemical Biology and Medicinal Chemistry, and Department of Pharmacology, University of North Carolina, Chapel Hill, NC 27599 (USA).
authors.forename: Robert M
authors.initials: RM
authors.lastname: Hughes
authors.forename: David S
authors.initials: DS
authors.lastname: Lawrence
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D000199:Actins / Q000378:metabolism; D019943:Amino Acid Substitution; D000818:Animals; D045744:Cell Line, Tumor; D002465:Cell Movement; D051338:Cofilin 1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D056931:Cryptochromes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008027:Light; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D051379:Mice; D018613:Microscopy, Confocal; D062308:Optogenetics; D011554:Pseudopodia / Q000378:metabolism; D000097573:Red Fluorescent Protein
chemical_list: D000199:Actins; D051338:Cofilin 1; D056931:Cryptochromes; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein
keywords: F-actin; cell motility; cofilin; optogenetics; protein design