A Light-Oxygen-Voltage Receptor Integrates Light and Temperature

Dietler, Julia; Schubert, Roman; Krafft, Tobias G.A.; Meiler, Simone; Kainrath, Stephanie; Richter, Florian; Schweimer, Kristian; Weyand, Michael; Janovjak, Harald; Möglich, Andreas

doi:10.1016/j.jmb.2021.167107

Cited by 21 publications

(28 citation statements)

References 85 publications

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“…We discovered that BcLOV4 is a temperature sensor in addition to its known role as a photosensor. While temperature-dependence has been observed in certain photosensors and optogenetic probes, this dependence mostly manifests as decreased protein stability or photoreactivity at elevated temperatures 3 , 10 , 35 - 39 . By contrast, BcLOV4 folds and translocates rapidly when exposed to light at all temperatures, but then, under sustained illumination, enters a long-lived inactive state and reverts to the cytoplasm at a rate that increases with both temperature and light dose ( Figure 2 , 3 , Supplementary Figure 10 ).…”

Section: Discussionmentioning

confidence: 99%

“…Such probes are typically engineered from proteins that evolved to respond to their host’s environmental conditions, i.e. to its light status 1 , 2 , but in some cases also to temperature 3 - 5 . Light-responsive actuators now exist for control of protein dimerization 6 - 10 , allostery 11 , 12 , oligomerization 13 , ion transport 14 , and membrane recruitment 15 , 16 , providing an extensive toolset for precise manipulation of an array of biological processes, including cell signaling.…”

Section: Introductionmentioning

confidence: 99%

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Temperature-responsive optogenetic probes of cell signaling

et al. 2021

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We describe single-component optogenetic probes whose activation dynamics depend on both light and temperature. We used the BcLOV4 photoreceptor to stimulate Ras and PI3K signaling in mammalian cells, allowing activation over a large dynamic range with low basal levels. Surprisingly, we found that BcLOV4 membrane translocation dynamics could be tuned by both light and temperature such that membrane localization spontaneously decayed at elevated temperatures despite constant illumination. Quantitative modeling predicted BcLOV4 activation dynamics across a range of light and temperature inputs and thus provides an experimental roadmap for BcLOV4-based probes. BcLOV4 drove strong and stable signal activation in both zebrafish and fly cells, and thermal inactivation provided a means to multiplex distinct blue-light sensitive tools in individual mammalian cells. BcLOV4 is thus a versatile photosensor with unique light and temperature sensitivity that enables straightforward generation of broadly applicable optogenetic tools.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Temperature-responsive optogenetic probes of cell signaling

et al. 2021

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“…As a corollary, the maximal difference between the two simulated photoreceptors which differ in their dark-recovery rates by a factor of ten is larger than in scenario C (dashed black line). (E) Certain photoreceptor circuits are sensitive to a second stimulus e.g., light of a different color, a chemical inducer, or changes in temperature (Dietler et al, 2021;Romano et al, 2021). Initial photostimulation (red bar) can be counteracted by subsequent application of the second signal (brown bar).…”

Section: Figurementioning

confidence: 99%

Light-regulated gene expression in Bacteria: Fundamentals, advances, and perspectives

Ohlendorf

Möglich²

2022

Front. Bioeng. Biotechnol.

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Numerous photoreceptors and genetic circuits emerged over the past two decades and now enable the light-dependent i.e., optogenetic, regulation of gene expression in bacteria. Prompted by light cues in the near-ultraviolet to near-infrared region of the electromagnetic spectrum, gene expression can be up- or downregulated stringently, reversibly, non-invasively, and with precision in space and time. Here, we survey the underlying principles, available options, and prominent examples of optogenetically regulated gene expression in bacteria. While transcription initiation and elongation remain most important for optogenetic intervention, other processes e.g., translation and downstream events, were also rendered light-dependent. The optogenetic control of bacterial expression predominantly employs but three fundamental strategies: light-sensitive two-component systems, oligomerization reactions, and second-messenger signaling. Certain optogenetic circuits moved beyond the proof-of-principle and stood the test of practice. They enable unprecedented applications in three major areas. First, light-dependent expression underpins novel concepts and strategies for enhanced yields in microbial production processes. Second, light-responsive bacteria can be optogenetically stimulated while residing within the bodies of animals, thus prompting the secretion of compounds that grant health benefits to the animal host. Third, optogenetics allows the generation of precisely structured, novel biomaterials. These applications jointly testify to the maturity of the optogenetic approach and serve as blueprints bound to inspire and template innovative use cases of light-regulated gene expression in bacteria. Researchers pursuing these lines can choose from an ever-growing, versatile, and efficient toolkit of optogenetic circuits.

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“…Prominent use cases for lightregulated gene expression in bacteria include the modulation of production processes 24,44 , the spatial patterning of expression and the templating of materials 36,37 , and the control of bacteria inside the digestive tract of animals 35,45 . The dominant strategies for achieving optogenetic control of bacterial gene expression employ either light-sensitive two-component systems [25][26][27]30,46 , protein-protein interactions [47][48][49] , or second-messenger signaling 50 . All these setups have in common that they operate at the DNA level and involve the regulation of transcription initiation.…”

Section: Discussionmentioning

confidence: 99%

Light-dependent Control of Bacterial Expression at the mRNA Level

Ranzani

Wehrmann

Kaiser

et al. 2022

Preprint

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Sensory photoreceptors mediate numerous light-dependent adaptations across organisms. In optogenetics, photoreceptors achieve the reversible, non-invasive, and spatiotemporally precise control by light of gene expression and other cellular processes. The light-oxygen-voltage receptor PAL binds to small RNA aptamers with sequence specificity upon blue-light illumination. By embedding the responsive aptamer in the ribosome-binding sequence of genes of interest, their expression can be downregulated by light. We developed the pCrepusculo and pAurora optogenetic systems that are based on PAL and allow to down- and upregulate, respectively, bacterial gene expression using blue light. Both systems are realized as compact, single plasmids that exhibit stringent blue-light responses with low basal activity and up to several ten-fold dynamic range. As PAL exerts light-dependent control at the RNA level, it can be combined with other optogenetic circuits that generally control transcription initiation. By integrating regulatory mechanisms operating at the DNA and mRNA levels, optogenetic circuits with emergent properties can thus be devised. As a case in point, the pEnumbra setup permits to upregulate gene expression under moderate blue light whereas strong blue light shuts off expression again. Beyond providing novel signal-responsive expression systems for diverse applications in biotechnology and synthetic biology, our work also illustrates how the light-dependent PAL-aptamer interaction can be harnessed for the control and interrogation of RNA-based processes.

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A Light-Oxygen-Voltage Receptor Integrates Light and Temperature

Cited by 21 publications

References 85 publications

Temperature-responsive optogenetic probes of cell signaling

Temperature-responsive optogenetic probes of cell signaling

Light-regulated gene expression in Bacteria: Fundamentals, advances, and perspectives

Light-dependent Control of Bacterial Expression at the mRNA Level

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