Shedding light on animal cryptochromes
| Autor: | Albrecht Berndt, Jean-Pierre Bouly, William Wu, Nathalie Hoang, Sylwia Kacprzak, Marie Picot, Margaret Ahmad, Eva Wolf, Robert Bittl, Erik Schleicher |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2008 |
| Předmět: |
Semiquinone
Circadian clock Gene Expression Biochemistry 0302 clinical medicine Cryptochrome Biology (General) Receptor 0303 health sciences biology General Neuroscience Drosophila melanogaster Synopsis Photoreceptor Cells Invertebrate Energy source General Agricultural and Biological Sciences Oxidation-Reduction Research Article Photoreceptor Cells Vertebrate Light Signal Transduction Ultraviolet Rays DNA repair QH301-705.5 Biophysics Flavoprotein Flavin group Protein degradation Spodoptera General Biochemistry Genetics and Molecular Biology Cell Line 03 medical and health sciences Biological Clocks Flavins Botany Animals Humans Circadian rhythm Eye Proteins Molecular Biology 030304 developmental biology Flavoproteins Organisms Genetically Modified General Immunology and Microbiology Electron Spin Resonance Spectroscopy Photoreceptor protein DNA photolyase biology.organism_classification Cryptochromes Cell culture biology.protein 030217 neurology & neurosurgery |
| Zdroj: | PLoS Biology, Vol 6, Iss 7, p e168 (2008) PLoS Biology, Vol 6, Iss 7, p e160 (2008) PLoS Biology |
| ISSN: | 1545-7885 1544-9173 |
| Popis: | Cryptochromes are a class of flavoprotein blue-light signaling receptors found in plants, animals, and humans that control plant development and the entrainment of circadian rhythms. In plant cryptochromes, light activation is proposed to result from photoreduction of a protein-bound flavin chromophore through intramolecular electron transfer. However, although similar in structure to plant cryptochromes, the light-response mechanism of animal cryptochromes remains entirely unknown. To complicate matters further, there is currently a debate on whether mammalian cryptochromes respond to light at all or are instead activated by non–light-dependent mechanisms. To resolve these questions, we have expressed both human and Drosophila cryptochrome proteins to high levels in living Sf21 insect cells using a baculovirus-derived expression system. Intact cells are irradiated with blue light, and the resulting cryptochrome photoconversion is monitored by fluorescence and electron paramagnetic resonance spectroscopic techniques. We demonstrate that light induces a change in the redox state of flavin bound to the receptor in both human and Drosophila cryptochromes. Photoreduction from oxidized flavin and subsequent accumulation of a semiquinone intermediate signaling state occurs by a conserved mechanism that has been previously identified for plant cryptochromes. These results provide the first evidence of how animal-type cryptochromes are activated by light in living cells. Furthermore, human cryptochrome is also shown to undergo this light response. Therefore, human cryptochromes in exposed peripheral and/or visual tissues may have novel light-sensing roles that remain to be elucidated. Author Summary Vision in animals is generally associated with light-sensitive rhodopsin pigments located in the eyes. However, animals ranging from flies to humans also possess ancient visual receptors known as cryptochromes in multiple cell types. In this work, we study the mechanism of light sensing in two representative animal cryptochromes: a light-sensitive Drosophila cryptochrome (Dmcry) and a presumed light-insensitive mammalian cryptochrome from humans (Hscry1). We expressed recombinant cryptochromes to high levels in living cells, irradiated the cells with blue light, and analyzed the proteins' response to irradiation with electron paramagnetic resonance and fluorescence spectroscopic techniques. Photoreduction of protein-bound oxidized FAD cofactor to its radical form emerged as the primary cryptochrome photoreaction in living cells, and was correlated with a light-sensitive biological response in whole organisms. These results indicate that both Dmcry and Hscry1 are capable of undergoing similar light-driven reactions and suggest the possibility of an as-yet unknown photo-perception role for human cryptochromes in tissues exposed to light. Cryptochromes are blue-light-absorbing receptors found in plants, animals, and humans. In mammals, they are not thought to respond to light, but this study demonstrates contrary evidence that indeed, human cryptochromes undergo a photochemical transformation in response to light. |
| Databáze: | OpenAIRE |
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