title: Control of protein function through optochemical translocation.
abstract: Controlled manipulation of proteins and their function is important in almost all biological disciplines. Here, we demonstrate control of protein activity with light. We present two different applications-light-triggered transcription and light-triggered protease cleavage-both based on the same concept of protein mislocation, followed by optochemically triggered translocation to an active cellular compartment. In our approach, we genetically encode a photocaged lysine into the nuclear localization signal (NLS) of the transcription factor SATB1. This blocks nuclear import of the protein until illumination induces caging group removal and release of the protein into the nucleus. In the first application, prepending this NLS to the transcription factor FOXO3 allows us to optochemically switch on its transcription activity. The second application uses the developed light-activated NLS to control nuclear import of TEV protease and subsequent cleavage of nuclear proteins containing TEV cleavage sites. The small size of the light-controlled NLS (only 20 amino acids) minimizes impact of its insertion on protein function and promises a general approach to a wide range of optochemical applications. Since the light-activated NLS is genetically encoded and optically triggered, it will prove useful to address a variety of problems requiring spatial and temporal control of protein function, for example, in stem-cell, developmental, and cancer biology.
authors.affiliation: Department of Chemistry and Center for NanoScience (CeNS), University of Munich (LMU) , Butenandtstraße 11, 81377 München, Germany.
authors.forename: Hanna
authors.initials: H
authors.lastname: Engelke
authors.forename: Chungjung
authors.initials: C
authors.lastname: Chou
authors.forename: Rajendra
authors.initials: R
authors.lastname: Uprety
authors.forename: Phillip
authors.initials: P
authors.lastname: Jess
authors.forename: Alexander
authors.initials: A
authors.lastname: Deiters
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000528:radiation effects; D000595:Amino Acid Sequence; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects; D000071316:Forkhead Box Protein O3; D051858:Forkhead Transcription Factors / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects; D057809:HEK293 Cells; D006801:Humans; D008027:Light; D036961:Matrix Attachment Region Binding Proteins / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects; D008969:Molecular Sequence Data; D019913:Nuclear Localization Signals / Q000235:genetics; D055668:Photochemical Processes; D015202:Protein Engineering / Q000379:methods*; D058615:Synthetic Biology
chemical_list: C499517:FOXO3 protein, human; D000071316:Forkhead Box Protein O3; D051858:Forkhead Transcription Factors; D036961:Matrix Attachment Region Binding Proteins; D019913:Nuclear Localization Signals; C076189:SATB1 protein, human; D010450:Endopeptidases; C099488:TEV protease
keywords: nuclear import; optogenetics; photocontrolled TEV-cleavage; photocontrolled transcription; protein control