Novel ATP-binding and autophosphorylation activity associated with Arabidopsis and human cryptochrome-1.
Cryptochromes are blue-light photoreceptors sharing sequence similarity to photolyases, a class of flavoenzymes catalyzing repair of UV-damaged DNA via electron transfer mechanisms. Despite significant amino acid sequence similarity in both catalytic and cofactor-binding domains, cryptochromes lack DNA repair functions associated with photolyases, and the molecular mechanism involved in cryptochrome signaling remains obscure. Here, we report a novel ATP binding and autophosphorylation activity associated with Arabidopsis cry1 protein purified from a baculovirus expression system. Autophosphorylation occurs on serine residue(s) and is absent in preparations of cryptochrome depleted in flavin and/or misfolded. Autophosphorylation is stimulated by light in vitro and oxidizing agents that act as flavin antagonists prevent this stimulation. Human cry1 expressed in baculovirus likewise shows ATP binding and autophosphorylation activity, suggesting this novel enzymatic activity may be important to the mechanism of action of both plant and animal cryptochromes.
Bouly
Jean-Pierre
JP
Université Paris VI, Paris, France.
Giovani
Baldissera
B
Djamei
Armin
A
Mueller
Markus
M
Zeugner
Anke
A
Dudkin
Elizabeth A
EA
Batschauer
Alfred
A
Ahmad
Margaret
M
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D017360:Arabidopsis / Q000378:metabolism*; D029681:Arabidopsis Proteins; D016367:Baculoviridae / Q000502:physiology; D001683:Biological Clocks; D001709:Biotechnology / Q000379:methods; D002460:Cell Line; D056931:Cryptochromes; D029721:Drosophila Proteins; D005136:Eye Proteins; D005415:Flavins / Q000037:antagonists & inhibitors / Q000378:metabolism; D005420:Flavoproteins / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D008027:Light; D009711:Nucleotides / Q000378:metabolism; D016877:Oxidants / Q000737:chemistry / Q000494:pharmacology; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation; D017956:Photoreceptor Cells, Invertebrate; D011485:Protein Binding / Q000502:physiology*; D043562:Receptors, G-Protein-Coupled; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012694:Serine / Q000378:metabolism; D018411:Spodoptera
D029681:Arabidopsis Proteins; C538962:CRY1 protein, Arabidopsis; C538963:CRY1 protein, human; D056931:Cryptochromes; D029721:Drosophila Proteins; D005136:Eye Proteins; D005415:Flavins; D005420:Flavoproteins; D009711:Nucleotides; D016877:Oxidants; D010761:Phosphorus Radioisotopes; D043562:Receptors, G-Protein-Coupled; D011994:Recombinant Proteins; C119828:cry protein, Drosophila; D012694:Serine; D000255:Adenosine Triphosphate