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.

[1]  Baldissera Giovani,et al.  Light-induced electron transfer in a cryptochrome blue-light photoreceptor , 2003, Nature Structural Biology.

[2]  T. Todo,et al.  Zebrafish CRY represses transcription mediated by CLOCK‐BMAL heterodimer without inhibiting its binding to DNA , 2002, Genes to cells : devoted to molecular & cellular mechanisms.

[3]  T. Mockler,et al.  Regulation of Arabidopsis cryptochrome 2 by blue-light-dependent phosphorylation , 2002, Nature.

[4]  Paul Galland,et al.  Action Spectrum for Cryptochrome-Dependent Hypocotyl Growth Inhibition in Arabidopsis1 , 2002, Plant Physiology.

[5]  Ralf Stanewsky,et al.  Clock mechanisms in Drosophila , 2002, Cell and Tissue Research.

[6]  E. Eide,et al.  The Circadian Regulatory Proteins BMAL1 and Cryptochromes Are Substrates of Casein Kinase Iε* , 2002, The Journal of Biological Chemistry.

[7]  O. Froy,et al.  Redox potential: differential roles in dCRY and mCRY1 functions. , 2002, Current biology : CB.

[8]  Haisun Zhu,et al.  A putative flavin electron transport pathway is differentially utilized in Xenopus CRY1 and CRY2 , 2001, Current Biology.

[9]  J. Hoch,et al.  PAS-A domain of phosphorelay sensor kinase A: A catalytic ATP-binding domain involved in the initiation of development in Bacillus subtilis , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[10]  A. Cashmore,et al.  The Signaling Mechanism of Arabidopsis CRY1 Involves Direct Interaction with COP1 Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.010367. , 2001, The Plant Cell Online.

[11]  S. Yokoyama,et al.  Crystal structure of thermostable DNA photolyase: Pyrimidine-dimer recognition mechanism , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[12]  T. Carell,et al.  The mechanism of action of DNA photolyases. , 2001, Current opinion in chemical biology.

[13]  Haiyang Wang,et al.  Direct Interaction of Arabidopsis Cryptochromes with COP1 in Light Control Development , 2001, Science.

[14]  C. Kyriacou,et al.  Light-dependent interaction between Drosophila CRY and the clock protein PER mediated by the carboxy terminus of CRY , 2001, Current Biology.

[15]  Yan Liu,et al.  The C Termini of Arabidopsis Cryptochromes Mediate a Constitutive Light Response , 2000, Cell.

[16]  J. Deisenhofer.  DNA photolyases and cryptochromes. , 2000, Mutation research.

[17]  A. Eker,et al.  Intraprotein radical transfer during photoactivation of DNA photolyase , 2000, Nature.

[18]  K Kume,et al.  Interacting molecular loops in the mammalian circadian clock. , 2000, Science.

[19]  T. Kondo,et al.  Nucleotide binding and autophosphorylation of the clock protein KaiC as a circadian timing process of cyanobacteria. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[20]  D. V. Leenen,et al.  Mammalian Cry1 and Cry2 are essential for maintenance of circadian rhythms , 1999, Nature.

[21]  J. C. Long,et al.  Involvement of Plasma Membrane Redox Activity and Calcium Homeostasis in the UV-B and UV-A/Blue Light Induction of Gene Expression in Arabidopsis , 1998, Plant Cell.

[22]  M. Ahmad,et al.  The CRY1 blue light photoreceptor of Arabidopsis interacts with phytochrome A in vitro. , 1998, Molecular cell.

[23]  T. Mockler,et al.  Enhancement of blue-light sensitivity of Arabidopsis seedlings by a blue light receptor cryptochrome 2. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[24]  S. Hardin,et al.  Low-voltage separation of phosphoamino acids by silica gel thin-layer electrophoresis in a DNA electrophoresis cell. , 1998, BioTechniques.

[25]  A. Cashmore,et al.  Chimeric Proteins between cry1 and cry2 Arabidopsis Blue Light Photoreceptors Indicate Overlapping Functions and Varying Protein Stability , 1998, Plant Cell.

[26]  Satoru Kanai,et al.  Molecular Evolution of the Photolyase–Blue-Light Photoreceptor Family , 1997, Journal of Molecular Evolution.

[27]  D. S. Hsu,et al.  Putative human blue-light photoreceptors hCRY1 and hCRY2 are flavoproteins. , 1996, Biochemistry.

[28]  P. Gueguen,et al.  ATP Binding and Hydrolysis by the Multifunctional Protein Disulfide Isomerase (*) , 1996, The Journal of Biological Chemistry.

[29]  M. Ahmad,et al.  Association of flavin adenine dinucleotide with the Arabidopsis blue light receptor CRY1 , 1995, Science.

[30]  J. Deisenhofer,et al.  Crystal structure of DNA photolyase from Escherichia coli. , 1995, Science.

[31]  A. Sancar,et al.  Putative blue-light photoreceptors from Arabidopsis thaliana and Sinapis alba with a high degree of sequence homology to DNA photolyase contain the two photolyase cofactors but lack DNA repair activity. , 1995, Biochemistry.

[32]  A. Cashmore,et al.  HY4 gene of A. thaliana encodes a protein with characteristics of a blue-light photoreceptor , 1993, Nature.

[33]  M. Porst,et al.  A PHOTORECEPTOR SYSTEM REGULATING in vivo AND in vitro PHOSPHORYLATION OF A PEA PLASMA MEMBRANE PROTEIN , 1984 .

[34]  Minoru Kanehisa,et al.  Identification of a new cryptochrome class. Structure, function, and evolution. , 2003, Molecular cell.

[35]  J. Knowles,et al.  [8] Photoaffinity labeling , 1977 .