title: Reaction mechanism of Drosophila cryptochrome.
abstract: Cryptochrome (CRY) is a blue-light sensitive flavoprotein that functions as the primary circadian photoreceptor in Drosophila melanogaster. The mechanism by which it transmits the light signal to the core clock circuitry is not known. We conducted in vitro studies on the light-induced conformational change in CRY and its effect on protein-protein interaction and performed in vivo analysis of the lifetime of the signaling state of the protein to gain some insight into the mechanism of phototransduction. We find that exposure of CRY to blue light induces a conformation similar to that of the constitutively active CRY mutant with a C-terminal deletion (CRYΔ). This light-induced conformation has a half-life of ∼15 min in the dark at 25 °C and is characterized by increased affinity to Jetlag E3 ligase. In vivo analysis reveals that in the Drosophila S2 cell line, the signaling state induced by a millisecond light exposure has a half-life of 27 min in the dark at 0 °C during which period it is susceptible to degradation by the ubiquitin-proteasome system. These findings lead to a plausible model for circadian photoreception/phototransduction in Drosophila.
authors.affiliation: Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, NC 27599, USA.
authors.forename: Nuri
authors.initials: N
authors.lastname: Ozturk
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Selby
authors.forename: Yunus
authors.initials: Y
authors.lastname: Annayev
authors.forename: Dongping
authors.initials: D
authors.lastname: Zhong
authors.forename: Aziz
authors.initials: A
authors.lastname: Sancar
mesh_terms: D000818:Animals; D002940:Circadian Rhythm; D056931:Cryptochromes / Q000737:chemistry*; D004331:Drosophila melanogaster / Q000378:metabolism*; D005420:Flavoproteins / Q000737:chemistry; D007700:Kinetics; D008027:Light; D008956:Models, Chemical; D009154:Mutation; D017956:Photoreceptor Cells, Invertebrate / Q000378:metabolism; D011487:Protein Conformation; D025941:Protein Interaction Mapping; D013696:Temperature; D013997:Time Factors
chemical_list: D056931:Cryptochromes; D005420:Flavoproteins