title: Blue light-dependent in vivo and in vitro phosphorylation of Arabidopsis cryptochrome 1.
abstract: Cryptochromes are photolyase-like blue/UV-A light receptors that regulate various light responses in animals and plants. Arabidopsis cryptochrome 1 (cry1) is the major photoreceptor mediating blue light inhibition of hypocotyl elongation. The initial photochemistry underlying cryptochrome function and regulation remain poorly understood. We report here a study of the blue light-dependent phosphorylation of Arabidopsis cry1. Cry1 is detected primarily as unphosphorylated protein in etiolated seedlings, but it is phosphorylated in plants exposed to blue light. Cry1 phosphorylation increases in response to increased fluence of blue light, whereas the phosphorylated cry1 disappears rapidly when plants are transferred from light to dark. Light-dependent cry1 phosphorylation appears specific to blue light, because little cry1 phosphorylation is detected in seedlings treated with red light or far-red light, and it is largely independent from phytochrome actions, because no phytochrome mutants tested significantly affect cry1 phosphorylation. The Arabidopsis cry1 protein expressed and purified from insect cells is phosphorylated in vitro in a blue light-dependent manner, consistent with cry1 undergoing autophosphorylation. To determine whether cry1 phosphorylation is associated with its function or regulation, we isolated and characterized missense cry1 mutants that express full-length CRY1 apoprotein. Mutant residues are found throughout the CRY1 coding sequence, but none of these inactive cry1 mutant proteins shows blue light-induced phosphorylation. These results demonstrate that blue light-dependent cry1 phosphorylation is closely associated with the function or regulation of the photoreceptor and that the overall structure of cry1 is critical to its phosphorylation.
authors.affiliation: Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles, California 90095, USA.
authors.forename: Dror
authors.initials: D
authors.lastname: Shalitin
authors.forename: Xuhong
authors.initials: X
authors.lastname: Yu
authors.forename: Maskit
authors.initials: M
authors.lastname: Maymon
authors.forename: Todd
authors.initials: T
authors.lastname: Mockler
authors.forename: Chentao
authors.initials: C
authors.lastname: Lin
mesh_terms: D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics / Q000528:radiation effects*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D056931:Cryptochromes; D029721:Drosophila Proteins; D005136:Eye Proteins; D005420:Flavoproteins / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D007700:Kinetics; D008027:Light; D008969:Molecular Sequence Data; D010766:Phosphorylation / Q000528:radiation effects; D017956:Photoreceptor Cells, Invertebrate; D043562:Receptors, G-Protein-Coupled; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
chemical_list: D029681:Arabidopsis Proteins; C538962:CRY1 protein, Arabidopsis; D056931:Cryptochromes; D029721:Drosophila Proteins; D005136:Eye Proteins; D005420:Flavoproteins; D043562:Receptors, G-Protein-Coupled; C119828:cry protein, Drosophila