No abstract

Living organisms detect changes in temperature using thermosensory molecules. However, these molecules and/or their mechanisms for sensing temperature differ among organisms. To identify thermosensory molecules in plants, we investigated chloroplast positioning in response to temperature changes and identified a blue-light photoreceptor, phototropin, that is an essential regulator of chloroplast positioning. Based on the biochemical properties of phototropin during the cellular response to light and temperature changes, we found that phototropin perceives temperature based on the temperature-dependent lifetime of the photoactivated chromophore. Our findings indicate that phototropin perceives both blue light and temperature and uses this information to arrange the chloroplasts for optimal photosynthesis. Because the photoactivated chromophore of many photoreceptors has a temperature-dependent lifetime, a similar temperature-sensing mechanism likely exists in other organisms. Thus, photoreceptors may have the potential to function as thermoreceptors.iving organisms perceive temperature using thermosensory molecules. Various types of thermosensory molecules have been identified in microorganisms, animals, and plants, and these molecules perceive temperature via temperature-dependent intraand intermolecular interactions (1-3). However, these thermosensory molecules and/or their mechanisms for temperature sensing differ among organisms (1).As photosynthetic organelles, chloroplasts change their intracellular position in response to light and temperature (4-7). For example, under low light conditions at 22°C, chloroplasts accumulate at the cell surface (along the periclinal cell wall) to maximize photosynthetic efficiency, in a phenomenon termed the accumulation response (4, 6). However, under the same light conditions, but at a temperature of 5°C, the chloroplasts move to the cell periphery (along the anticlinal cell wall), possibly to avoid the light, in a phenomenon termed the cold-avoidance response or the cold-positioning response (5, 6). Because the cold-avoidance response is temperature dependent (5), the response is likely controlled by a thermosensory molecule. In a previous study, we found that the blue-light (BL) photoreceptor phototropin (phot) is responsible for the cold-avoidance response in the fern Adiantum capillus-veneris, which has three types of phot molecules (phot1, phot2, and neochrome1) (5,8). In this fern, phot2 mediates the cold-avoidance response (5). Therefore, in the present study, we hypothesized that phot is involved in temperature perception and we further analyzed the cold-avoidance response in the liverwort Marchantia polymorpha, which has only a single copy of the PHOT gene (MpPHOT) (9). Results and DiscussionFirst, we examined whether MpPHOT is essential for the coldavoidance response in M. polymorpha. When the wild type (WT), the knockout mutant (Mpphot KO ), and the complementation line (Mpphot/Mpphot KO ) (9) (Fig. S1) were treated with low white light at 5°C for 24 h to induce ...

BLUF (a sensor of Blue-Light Using FAD) is a novel putative photoreceptor domain that is found in many bacteria and some eukaryotic algae. As found on genome analysis, certain cyanobacteria have BLUF proteins with a short C-terminal extension. As typical examples, Tll0078 from thermophilic Thermosynechococcus elongatus BP-1 and Slr1694 from mesophilic Synechocystis sp. PCC 6803 were comparatively studied. FAD of both proteins was hardly reduced by exogenous reductants or mediators except methylviologen but showed a typical spectral shift to a longer wavelength upon excitation with blue light. In particular, freshly prepared Tll0078 protein showed slow but reversible aggregation, indicative of light-induced conformational changes in the protein structure. Tll0078 is far more stable as to heat treatment than Slr1694, as judged from flavin fluorescence. The slr1694-disruptant showed phototactic motility away from the light source (negative phototaxis), while the wild type Synechocystis showed positive phototaxis toward the source. Yeast two-hybrid screening with slr1694 showed self-interaction of Slr1694 (PixD) with itself and interaction with a novel PatA-like response regulator, Slr1693 (PixE). These results were discussed in relation to the signaling mechanism of the "short" BLUF proteins in the regulation of cyanobacterial phototaxis.

Proteins with a BLUF (sensor of blue light using flavin adenine dinucleotide) domain represent a newly recognized class of photoreceptors that is widely distributed in the genomes of photosynthetic bacteria, cyanobacteria, and Euglena. Recently, Okajima et al. [Okajima, K., Yoshihara, S., Geng, X., Katayama, M. and Ikeuchi, M. (2003) Plant Cell Physiol. 44 (Suppl), 162] purified BLUF protein Tll0078 encoded in the genome of thermophilic cyanobacterium Thermosynechococcus elongatus BP-1 by expressing the protein in Escherichia coli. We investigated the photocycle of Tll0078 by measuring the picosecond fluorescence kinetics, transient absorption changes, and the UV-visible absorption spectra at 10 to 330 K. The absorption spectrum of the FAD moiety of Tll0078 showed a 10-nm red shift upon illumination at 278-330 K. The quantum efficiency of the formation of the red-shifted form was 29%. Illumination at 10 K, on the other hand, caused only a 5-nm red shift in about one-half of the protein population. The 5-nm-shifted form was stable at 10 K. The 5-nm red-shifted form was converted into the 10-nm red-shifted form at 50-240 K upon warming in the dark. At room temperature, the 10-nm red-shifted final product appeared within 10 ns after laser flash excitation. The lifetime of the fluorescence of FAD was found to be 120 ps at room temperature. These results reveal a fast and efficient photoconversion process from the singlet-excited state to the final product at room temperature. A photocycle of BLUF protein is proposed that includes the 5-nm red-shifted intermediate form as the precursor for the 10-nm red-shifted final product. The temperature dependence of each step of the photocycle is also discussed.