Arthaut, Louis-David, Jourdan, Nathalie, Mteyrek, Ali, Procopio, Maria ORCID: 0000-0002-2521-3067, El-Esawi, Mohamed, d'Harlingue, Alain, Bouchet, Pierre-Etienne, Witczak, Jacques, Ritz, Thorsten ORCID: 0000-0002-2713-539X, Klarsfeld, Andre ORCID: 0000-0001-9293-4887, Birman, Serge, Usselman, Robert J., Hoecker, Ute ORCID: 0000-0002-5636-9777, Martino, Carlos F. and Ahmad, Margaret (2017). Blue-light induced accumulation of reactive oxygen species is a consequence of the Drosophila cryptochrome photocycle. PLoS One, 12 (3). SAN FRANCISCO: PUBLIC LIBRARY SCIENCE. ISSN 1932-6203

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Abstract

Cryptochromes are evolutionarily conserved blue-light absorbing flavoproteins which participate in many important cellular processes including in entrainment of the circadian clock in plants, Drosophila and humans. Drosophila melanogaster cryptochrome (DmCry) absorbs light through a flavin (FAD) cofactor that undergoes photoreduction to the anionic radical (FAD(center dot-)) redox state both in vitro and in vivo. However, recent efforts to link this photoconversion to the initiation of a biological response have remained controversial. Here, we show by kinetic modeling of the DmCry photocycle that the fluence dependence, quantum yield, and half-life of flavin redox state interconversion are consistent with the anionic radical (FAD(center dot-)) as the signaling state in vivo. We show by fluorescence detection techniques that illumination of purified DmCry results in enzymatic conversion of molecular oxygen (O-2) to reactive oxygen species (ROS). We extend these observations in living cells to demonstrate transient formation of superoxide (O-2(center dot-)), and accumulation of hydrogen peroxide (H2O2) in the nucleus of insect cell cultures upon DmCry illumination. These results define the kinetic parameters of the Drosophila cryptochrome photocycle and support light-driven electron transfer to the flavin in DmCry signaling. They furthermore raise the intriguing possibility that light-dependent formation of ROS as a byproduct of the cryptochrome photocycle may contribute to its signaling role.

Item Type: Journal Article
Creators:
CreatorsEmailORCIDORCID Put Code
Arthaut, Louis-DavidUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Jourdan, NathalieUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Mteyrek, AliUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Procopio, MariaUNSPECIFIEDorcid.org/0000-0002-2521-3067UNSPECIFIED
El-Esawi, MohamedUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
d'Harlingue, AlainUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Bouchet, Pierre-EtienneUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Witczak, JacquesUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Ritz, ThorstenUNSPECIFIEDorcid.org/0000-0002-2713-539XUNSPECIFIED
Klarsfeld, AndreUNSPECIFIEDorcid.org/0000-0001-9293-4887UNSPECIFIED
Birman, SergeUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Usselman, Robert J.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Hoecker, UteUNSPECIFIEDorcid.org/0000-0002-5636-9777UNSPECIFIED
Martino, Carlos F.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Ahmad, MargaretUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
URN: urn:nbn:de:hbz:38-236602
DOI: 10.1371/journal.pone.0171836
Journal or Publication Title: PLoS One
Volume: 12
Number: 3
Date: 2017
Publisher: PUBLIC LIBRARY SCIENCE
Place of Publication: SAN FRANCISCO
ISSN: 1932-6203
Language: English
Faculty: Faculty of Mathematics and Natural Sciences
Divisions: Faculty of Mathematics and Natural Sciences > Department of Biology > Botanical Institute
Subjects: no entry
Uncontrolled Keywords:
KeywordsLanguage
ARABIDOPSIS CRYPTOCHROME; MOLECULAR-OXYGEN; DNA PHOTOLYASE; FLAVIN; MECHANISM; PHOTORECEPTORS; PATHWAYS; RHYTHMS; SENSOR; STATESMultiple languages
Multidisciplinary SciencesMultiple languages
Refereed: Yes
URI: http://kups.ub.uni-koeln.de/id/eprint/23660

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