title: Controlling Protein Activity and Degradation Using Blue Light.
abstract: Regulation of protein stability is a fundamental process in eukaryotic cells and pivotal to, e.g., cell cycle progression, faithful chromosome segregation, or protein quality control. Synthetic regulation of protein stability requires conditional degradation sequences (degrons) that induce a stability switch upon a specific signal. Fusion to a selected target protein permits to influence virtually every process in a cell. Light as signal is advantageous due to its precise applicability in time, space, quality, and quantity. Light control of protein stability was achieved by fusing the LOV2 photoreceptor domain of Arabidopsis thaliana phototropin1 with a synthetic degron (cODC1) derived from the carboxy-terminal degron of ornithine decarboxylase to obtain the photosensitive degron (psd) module. The psd module can be attached to the carboxy terminus of target proteins that are localized to the cytosol or nucleus to obtain light control over their stability. Blue light induces structural changes in the LOV2 domain, which in turn lead to activation of the degron and thus proteasomal degradation of the whole fusion protein. Variants of the psd module with diverse characteristics are useful to fine-tune the stability of a selected target at permissive (darkness) and restrictive conditions (blue light).
authors.affiliation: Department of Biology/Genetics, Philipps-University Marburg, Karl-von-Frisch-Str. 8, 35043, Marburg, Germany.
authors.forename: Anne P
authors.initials: AP
authors.lastname: Lutz
authors.affiliation: Department of Biology/Genetics, Philipps-University Marburg, Karl-von-Frisch-Str. 8, 35043, Marburg, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Renicke
authors.affiliation: Department of Biology/Genetics, Philipps-University Marburg, Karl-von-Frisch-Str. 8, 35043, Marburg, Germany. taxis@biologie.uni-marburg.de.
authors.forename: Christof
authors.initials: C
authors.lastname: Taxis
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017360:Arabidopsis / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008027:Light; D051379:Mice; D062308:Optogenetics / Q000379:methods*; D009955:Ornithine Decarboxylase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D010750:Phosphoproteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D018462:Photobiology / Q000379:methods; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D000072417:Protein Domains / Q000528:radiation effects; D017346:Protein Serine-Threonine Kinases; D055550:Protein Stability / Q000528:radiation effects; D059748:Proteolysis / Q000528:radiation effects; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects; D058615:Synthetic Biology / Q000379:methods; D014170:Transformation, Genetic
chemical_list: D029681:Arabidopsis Proteins; D004268:DNA-Binding Proteins; C000612094:LOV2 protein, Arabidopsis; D010750:Phosphoproteins; D011993:Recombinant Fusion Proteins; C103646:NPH1 protein, Arabidopsis; D017346:Protein Serine-Threonine Kinases; D046988:Proteasome Endopeptidase Complex; D009955:Ornithine Decarboxylase
keywords: Blue light; Degron; LOV2 domain; Optogenetics; Proteasome; Protein degradation; Protein stability; Synthetic biology; Ubiquitin-independent degradation