title: Genetically engineered photoinducible homodimerization system with improved dimer-forming efficiency.
abstract: Vivid (VVD) is a photoreceptor derived from Neurospora Crassa that rapidly forms a homodimer in response to blue light. Although VVD has several advantages over other photoreceptors as photoinducible homodimerization system, VVD has a critical limitation in its low dimer-forming efficiency. To overcome this limitation of wild-type VVD, here we conduct site-directed saturation mutagenesis in the homodimer interface of VVD. We have found that the Ile52Cys mutation of VVD (VVD-52C) substantially improves its homodimer-forming efficiency up to 180%. We have demonstrated the utility of VVD-52C for making a light-inducible gene expression system more robust. In addition, using VVD-52C, we have developed photoactivatable caspase-9, which enables optical control of apoptosis of mammalian cells. The present genetically engineered photoinducible homodimerization system can provide a powerful tool to optically control a broad range of molecular processes in the cell.
authors.affiliation: Graduate School of Arts and Sciences, The University of Tokyo , Komaba, Meguro-ku, Tokyo 153-8902, Japan.
authors.forename: Yuta
authors.initials: Y
authors.lastname: Nihongaki
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Suzuki
authors.forename: Fuun
authors.initials: F
authors.lastname: Kawano
authors.forename: Moritoshi
authors.initials: M
authors.lastname: Sato
mesh_terms: D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000528:radiation effects; D015966:Gene Expression Regulation, Fungal / Q000528:radiation effects; D008027:Light; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D009492:Neurospora crassa / Q000378:metabolism / Q000528:radiation effects; D062308:Optogenetics / Q000379:methods*; D015202:Protein Engineering; D055503:Protein Multimerization; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000528:radiation effects
chemical_list: D005656:Fungal Proteins; D011993:Recombinant Fusion Proteins; C423283:VVD protein, Neurospora crassa