• Journal of Photoscience
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• v.9 no.2
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• pp.335-337
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• 2002

Blue light (BL) induces stomatal opening through activation of H$^{+}$ pump, which creates electrical gradient across the plasma membrane for $K^{+}$ uptake into guard cells. The pump is the plasma membrane H$^{+}$ -ATPase and is activated via phosphorylation of the C-terminus with concomitant binding of the 14-3-3 protein. The opening is initiated by the perception of BL through phototropin (phot), which are recently identified as BL receptors in stomatal guard cells. In this study, we provide the biochemical evidence for phots as BL receptors in stomatal guard cells. vfphot was phosphorylated reversibly by BL, and phosphorylation levels of vfphot increased earlier than those of the plasma membrane W-ATPase. BL-dependent phosphorylations of vfphot and H$^{+}$-ATPase showed similar fluence dependency. Staurosporin, an inhibitor of serine/threonine protein kinase, and diphenyleneiodonium chloride (DPI), an inhibitor of flavoprotein, inhibited BL-dependent phosphorylations of vfphot and H$^{+}$ -ATPase. These results indicate that vfphot acts as a BL-receptor mediating stomatal opening.l opening.

• ALGAE
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• v.32 no.3
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• pp.235-244
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• 2017

Freshwater algae living in shallow waters have evolved various photomovement to stay in the optimum light condition for survival. Previous action-spectra investigations showed that Spirogyra filaments have phototropic movement in blue light. To decipher the genetic control of phototropic movement, two phototropin homologues were isolated from Spirogyra varians, and named SvphotA and SvphotB. Both phototropins have similar molecular structure consisted of two light-oxygen-voltage domains (LOV1, LOV2) and a serine / threonine kinase domain. SvphotA and SvphotB had 48.7% sequence identity. Phylogenetic analysis showed SvphotA and SvphotB belong to different clades suggesting early divergence, possibly before the divergence of land plants from the Zygnematales. Quantitative PCR and northern blot analysis showed that SvphotA and SvphotB responded differently to red and blue light. SvphotA was consistently expressed in the dark and in blue light, while SvphotB was expressed only when the plants were exposed to light. When the filaments were exposed to red light, SvphotA was significantly downregulated whereas SvphotB was highly upregulated. These results suggest that the two phototropins may have different roles in the photoresponse in S. varians.

• Proceedings of the Microbiological Society of Korea Conference
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• 2005.05a
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• pp.64-66
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• 2005

Photosynthetic microbes possess a wealth of photoactive proteins including chlorophyll-based pigments, phototropin-related blue light receptors, phytochromes, and cryptochromes. Surprisingly, recent genome sequencing projects discovered additional photoactive proteins, retinal-based rhodopsins, in cyanobacterial and algal genera. Most of these newly found rhodopsin genes and retinal synthase have not been expressed and their functions are unknown. Analysis of the Anabaena and Chlamyrhodopsin with retinal synthase revealed that they have sensory functions, which, based on our work with haloarchaeal rhodopsins, may use a variety of signaling mechanisms. Anabaena rhodopsin is believed to be sensory, shown to interact with a soluble transducer and the putative function is either chromatic adaptation or circadian rhythm. Chlamydomonas rhodopsins are involved in phototaxis and photophobic responses based on electrical measurements by RNAi experiment. In order to analyze the protein, we developed a sensory rhodopsin expression system in E. coli. The opsin in E. coil bound endogenous all-trans retinal to form a pigment and can be observed on the plate. Using this system we could identify retinal synthase in Anabaena PCC 7120. We conclude that Anabaena D475 dioxygenase functions as a retinal synthase to the Anabaena rhodopsin in the cell.