title: An optimized optogenetic clustering tool for probing protein interaction and function.
abstract: The Arabidopsis photoreceptor cryptochrome 2 (CRY2) was previously used as an optogenetic module, allowing spatiotemporal control of cellular processes with light. Here we report the development of a new CRY2-derived optogenetic module, 'CRY2olig', which induces rapid, robust, and reversible protein oligomerization in response to light. Using this module, we developed a novel protein interaction assay, Light-Induced Co-clustering, that can be used to interrogate protein interaction dynamics in live cells. In addition to use probing protein interactions, CRY2olig can also be used to induce and reversibly control diverse cellular processes with spatial and temporal resolution. Here we demonstrate disrupting clathrin-mediated endocytosis and promoting Arp2/3-mediated actin polymerization with light. These new CRY2-based approaches expand the growing arsenal of optogenetic strategies to probe cellular function.
authors.affiliation: Department of Pharmacology, University of Colorado School of Medicine, Aurora, Colorado 80045, USA.
authors.forename: Amir
authors.initials: A
authors.lastname: Taslimi
authors.affiliation: Department of Pharmacology, University of Colorado School of Medicine, Aurora, Colorado 80045, USA.
authors.forename: Justin D
authors.initials: JD
authors.lastname: Vrana
authors.affiliation: Department of Pharmacology, University of Colorado School of Medicine, Aurora, Colorado 80045, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Chen
authors.affiliation: Raymond and Beverly Sackler Laboratory of Genetics and Molecular Medicine, Department of Genetics and Molecular Biology, University of Connecticut Health Center, Farmington, Connecticut 06030, USA.
authors.forename: Sofya
authors.initials: S
authors.lastname: Borinskaya
authors.affiliation: Raymond and Beverly Sackler Laboratory of Genetics and Molecular Medicine, Department of Genetics and Molecular Biology, University of Connecticut Health Center, Farmington, Connecticut 06030, USA.
authors.forename: Bruce J
authors.initials: BJ
authors.lastname: Mayer
authors.affiliation: Department of Pharmacology, University of Colorado School of Medicine, Aurora, Colorado 80045, USA.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Kennedy
authors.affiliation: Department of Pharmacology, University of Colorado School of Medicine, Aurora, Colorado 80045, USA.
authors.forename: Chandra L
authors.initials: CL
authors.lastname: Tucker
mesh_terms: D051377:Actin-Related Protein 2 / Q000378:metabolism; D000199:Actins / Q000737:chemistry; D000818:Animals; D017360:Arabidopsis / Q000235:genetics; D019556:COS Cells; D002467:Cell Nucleus / Q000378:metabolism; D002522:Chlorocebus aethiops; D002966:Clathrin / Q000737:chemistry; D016000:Cluster Analysis; D003599:Cytoskeleton / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D004705:Endocytosis; D057809:HEK293 Cells; D006801:Humans; D008027:Light; D009154:Mutation; D062308:Optogenetics / Q000379:methods*; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D020798:Two-Hybrid System Techniques
chemical_list: D051377:Actin-Related Protein 2; D000199:Actins; D002966:Clathrin