The inherent antibiotic activity of myxobacteria-derived autofluorescent outer membrane vesicles is switched on and off by light stimulation

Lapuhs, Philipp; Schulz, Eilien; Garcia, Ronald; Goes, Adriely; Frank, Nicolas; Bollenbach, Lukas; Stibane, Veronika; Kuhn, Thomas; Koch, Marcus; Kiemer, Alexandra K.; Müller, Rolf; Fuhrmann, Kathrin; Fuhrmann, Gregor

doi:10.1039/d2nr02743g

Cited by 3 publications

(4 citation statements)

References 37 publications

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“…Our results provide detailed structural insight into the myxobacterial two-component signaling pathway, featuring light as an environmental signal in one of the most complex prokaryotic organisms with a unique life cycle and abundant synthesis of secondary metabolites with potential antimicrobial and anticancer properties (Lapuhs, Heinrich et al 2022). The described cryo-EM structures explain over half a century of structural research on plant and bacterial phytochromes limited by availability of crystals of intact photoreceptors.…”

Section: Discussionmentioning

confidence: 99%

“…In S. aurantica fruiting body formation is controlled by red and far-red light, implicating bacteriophytochrome (BphP) signaling (Qualls, Stephens et al 1978, White, Shropshire et al 1980, Woitowich, Halavaty et al 2018). Myxobacteria are also extensively studied for the synthesis of secondary metabolites with anticancer and antimicrobial properties, recently discovered to be modified by light (Schaberle, Lohr et al 2014, Lapuhs, Heinrich et al 2022).…”

Section: Introductionmentioning

confidence: 99%

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Signal Transduction in an Enzymatic Photoreceptor Revealed by Cryo-Electron Microscopy

Malla,

Hernandez,

Menendez

et al. 2023

Preprint

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Phytochromes are essential photoreceptor proteins in plants with homologs in bacteria and fungi that regulate a variety of important environmental responses. They display a reversible photocycle between two distinct states, the red-light absorbing Pr and the far-red light absorbing Pfr, each with its own structure. The reversible Pr to Pfr photoconversion requires covalently bound bilin chromophore and regulates the activity of a C-terminal enzymatic domain, which is usually a histidine kinase (HK). In plants, phytochromes translocate to nucleus where the C-terminal effector domain interacts with protein interaction factors (PIFs) to induce gene expression. In bacteria, the HK phosphorylates a response-regulator (RR) protein triggering downstream gene expression through a two-component signaling pathway. Although plant and bacterial phytochromes share similar structural composition, they have contrasting activity in the presence of light with most BphPs being active in the dark. The molecular mechanism that explains bacterial and plant phytochrome signaling has not been well understood due to limited structures of full-length phytochromes with enzymatic domain resolved at or near atomic resolution in both Pr and Pfr states. Here, we report the first Cryo-EM structures of a wild-type bacterial phytochrome with a HK enzymatic domain, determined in both Pr and Pfr states, between 3.75 and 4.13 Å resolution, respectively. Furthermore, we capture a distinct Pr/Pfr heterodimer of the same protein as potential signal transduction intermediate at 3.75 Å resolution. Our three Cryo-EM structures of the distinct signaling states of BphPs are further reinforced by Cryo-EM structures of the truncated PCM of the same protein determined for the Pr/Pfr heterodimer as well as Pfr state. These structures provide insight into the different light-signaling mechanisms that could explain how bacteria and plants see the light.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Signal Transduction in an Enzymatic Photoreceptor Revealed by Cryo-Electron Microscopy

Malla,

Hernandez,

Menendez

et al. 2023

Preprint

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“…Live bacteria possess a variety of endogenous fluorescent biological compounds with specific intrinsic excitation and emission wavelengths that can be used in applications such as label-free probes for bacterial detection and characterization purposes, for monitoring bacterial metabolic functions [ 1 ], and as a tracking mechanism of outer-membrane carrier vesicles [ 2 ], to mention a few. One microbial community of interest is the Streptomyces bacterial community.…”

Section: Introductionmentioning

confidence: 99%

Simultaneous Two- and Three-Photon Deep Imaging of Autofluorescence in Bacterial Communities

Fernández,

Classen,

Josyula

et al. 2024

Sensors

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The intrinsic fluorescence of bacterial samples has a proven potential for label-free bacterial characterization, monitoring bacterial metabolic functions, and as a mechanism for tracking the transport of relevant components through vesicles. The reduced scattering and axial confinement of the excitation offered by multiphoton imaging can be used to overcome some of the limitations of single-photon excitation (e.g., scattering and out-of-plane photobleaching) to the imaging of bacterial communities. In this work, we demonstrate in vivo multi-photon microscopy imaging of Streptomyces bacterial communities, based on the excitation of blue endogenous fluorophores, using an ultrafast Yb-fiber laser amplifier. Its parameters, such as the pulse energy, duration, wavelength, and repetition rate, enable in vivo multicolor imaging with a single source through the simultaneous two- and three-photon excitation of different fluorophores. Three-photon excitation at 1040 nm allows fluorophores with blue and green emission spectra to be addressed (and their corresponding ultraviolet and blue single-photon excitation wavelengths, respectively), and two-photon excitation at the same wavelength allows fluorophores with yellow, orange, or red emission spectra to be addressed (and their corresponding green, yellow, and orange single-photon excitation wavelengths). We demonstrate that three-photon excitation allows imaging over a depth range of more than 6 effective attenuation lengths to take place, corresponding to an 800 micrometer depth of imaging, in samples with a high density of fluorescent structures.

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“…In S. aurantiaca, fruiting body formation is controlled by red and far-red light, implicating bacteriophytochrome (BphP) signaling (13,16,17). Myxobacteria are also extensively studied for the synthesis of secondary metabolites with potential anticancer and antimicrobial properties, recently discovered to be modified by light (18,19).…”

Section: Introductionmentioning

confidence: 99%

Photoreception and signaling in bacterial phytochrome revealed by single-particle cryo-EM

Malla,

Hernandez,

Muniyappan

et al. 2024

Sci. Adv.

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Phytochromes are red-light photoreceptors discovered in plants with homologs in bacteria and fungi that regulate a variety of physiological responses. They display a reversible photocycle between two distinct states: a red-light–absorbing Pr state and a far-red light–absorbing Pfr state. The photoconversion regulates the activity of an enzymatic domain, usually a histidine kinase (HK). The molecular mechanism that explains how light controls the HK activity is not understood because structures of unmodified bacterial phytochromes with HK activity are missing. Here, we report three cryo–electron microscopy structures of a wild-type bacterial phytochrome with HK activity determined as Pr and Pfr homodimers and as a Pr/Pfr heterodimer with individual subunits in distinct states. We propose that the Pr/Pfr heterodimer is a physiologically relevant signal transduction intermediate. Our results offer insight into the molecular mechanism that controls the enzymatic activity of the HK as part of a bacterial two-component system that perceives and transduces light signals.

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The inherent antibiotic activity of myxobacteria-derived autofluorescent outer membrane vesicles is switched on and off by light stimulation

Abstract: Outer membrane vesicles are small, lipid-based vesicles shed from the outer membrane of Gram-negative bacteria. They are becoming increasingly recognised as important factors for resistance gene transfer, bacterial virulence factors...

Cited by 3 publications

References 37 publications

Signal Transduction in an Enzymatic Photoreceptor Revealed by Cryo-Electron Microscopy

Signal Transduction in an Enzymatic Photoreceptor Revealed by Cryo-Electron Microscopy

Simultaneous Two- and Three-Photon Deep Imaging of Autofluorescence in Bacterial Communities

Photoreception and signaling in bacterial phytochrome revealed by single-particle cryo-EM

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