The rhodopsin-guanylyl cyclase of the aquatic fungus Blastocladiella emersonii enables fast optical control of cGMP signaling.
Blastocladiomycota fungi form motile zoospores that are guided by sensory photoreceptors to areas of optimal light conditions. We showed that the microbial rhodopsin of Blastocladiella emersonii is a rhodopsin-guanylyl cyclase (RhGC), a member of a previously uncharacterized rhodopsin class of light-activated enzymes that generate the second messenger cyclic guanosine monophosphate (cGMP). Upon application of a short light flash, recombinant RhGC converted within 8 ms into a signaling state with blue-shifted absorption from which the dark state recovered within 100 ms. When expressed in Xenopus oocytes, Chinese hamster ovary cells, or mammalian neurons, RhGC generated cGMP in response to green light in a light dose-dependent manner on a subsecond time scale. Thus, we propose RhGC as a versatile tool for the optogenetic analysis of cGMP-dependent signaling processes in cell biology and the neurosciences.
Institute for Biology, Experimental Biophysics, Humboldt-Universität zu Berlin, D-10115 Berlin, Germany.
Ulrike
U
Scheib
Institute for Biology, Experimental Biophysics, Humboldt-Universität zu Berlin, D-10115 Berlin, Germany.
Katja
K
Stehfest
Institute for Synaptic Physiology, Center for Molecular Neurobiology Hamburg, University Medical Center Hamburg-Eppendorf, D-20251 Hamburg, Germany.
Christine E
CE
Gee
Department of Molecular Sensory Systems, Center of Advanced European Studies and Research (caesar), Ludwig-Erhard-Allee 2, 53175 Bonn, Germany.
Heinz G
HG
Körschen
Institute for Biology, Experimental Biophysics, Humboldt-Universität zu Berlin, D-10115 Berlin, Germany.
Roman
R
Fudim
Institute for Synaptic Physiology, Center for Molecular Neurobiology Hamburg, University Medical Center Hamburg-Eppendorf, D-20251 Hamburg, Germany. hegemann@rz.hu-berlin.de thomas.oertner@zmnh.uni-hamburg.de.
Thomas G
TG
Oertner
Institute for Biology, Experimental Biophysics, Humboldt-Universität zu Berlin, D-10115 Berlin, Germany. hegemann@rz.hu-berlin.de thomas.oertner@zmnh.uni-hamburg.de.
Peter
P
Hegemann
D000818:Animals; D001754:Blastocladiella / Q000201:enzymology* / Q000235:genetics; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D006152:Cyclic GMP / Q000235:genetics / Q000378:metabolism*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D006162:Guanylate Cyclase / Q000235:genetics / Q000378:metabolism*; D012243:Rhodopsin / Q000235:genetics / Q000378:metabolism; D015290:Second Messenger Systems / Q000502:physiology*; D014982:Xenopus laevis
D005656:Fungal Proteins; D012243:Rhodopsin; D006162:Guanylate Cyclase; D006152:Cyclic GMP