title: Structures of Drosophila cryptochrome and mouse cryptochrome1 provide insight into circadian function.
abstract: Drosophila cryptochrome (dCRY) is a FAD-dependent circadian photoreceptor, whereas mammalian cryptochromes (CRY1/2) are integral clock components that repress mCLOCK/mBMAL1-dependent transcription. We report crystal structures of full-length dCRY, a dCRY loop deletion construct, and the photolyase homology region of mouse CRY1 (mCRY1). Our dCRY structures depict Phe534 of the regulatory tail in the same location as the photolesion in DNA-repairing photolyases and reveal that the sulfur loop and tail residue Cys523 plays key roles in the dCRY photoreaction. Our mCRY1 structure visualizes previously characterized mutations, an NLS, and MAPK and AMPK phosphorylation sites. We show that the FAD and antenna chromophore-binding regions, a predicted coiled-coil helix, the C-terminal lid, and charged surfaces are involved in FAD-independent mPER2 and FBXL3 binding and mCLOCK/mBMAL1 transcriptional repression. The structure of a mammalian cryptochrome1 protein may catalyze the development of CRY chemical probes and the design of therapeutic metabolic modulators.
authors.affiliation: Department of Physiological Chemistry and Centre for Integrated Protein Science Munich (CIPSM), Butenandt Institute, Ludwig Maximilians University of Munich, Butenandtstrasse 5, 81377 Munich, Germany.
authors.forename: Anna
authors.initials: A
authors.lastname: Czarna
authors.forename: Alex
authors.initials: A
authors.lastname: Berndt
authors.forename: Hari Raj
authors.initials: HR
authors.lastname: Singh
authors.forename: Astrid
authors.initials: A
authors.lastname: Grudziecki
authors.forename: Andreas G
authors.initials: AG
authors.lastname: Ladurner
authors.forename: Gyula
authors.initials: G
authors.lastname: Timinszky
authors.forename: Achim
authors.initials: A
authors.lastname: Kramer
authors.forename: Eva
authors.initials: E
authors.lastname: Wolf
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D057906:Circadian Clocks; D002940:Circadian Rhythm; D056931:Cryptochromes / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004252:DNA Mutational Analysis; D004330:Drosophila / Q000378:metabolism*; D029721:Drosophila Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004579:Electron Transport; D005136:Eye Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D044783:F-Box Proteins / Q000378:metabolism; D005786:Gene Expression Regulation; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D056950:Period Circadian Proteins / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D016415:Sequence Alignment; D014158:Transcription, Genetic
chemical_list: C538964:Cry1 protein, mouse; D056931:Cryptochromes; D029721:Drosophila Proteins; D005136:Eye Proteins; D044783:F-Box Proteins; C519794:Fbxl3 protein, mouse; C513312:Per2 protein, mouse; D056950:Period Circadian Proteins; C119828:cry protein, Drosophila; C529384:Parp9 protein, mouse; D011065:Poly(ADP-ribose) Polymerases