Optogenetic control of integrin-matrix interaction

Baaske, Julia; Mühlhäuser, Wignand W. D.; Yousefi, O. Sascha; Zanner, Sebastian; Radziwill, Gerald; Hörner, Maximilian; Schamel, Wolfgang W. A.; Weber, Wilfried

doi:10.1038/s42003-018-0264-7

Cited by 32 publications

(34 citation statements)

References 38 publications

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“…The light-responsive N-terminal 651 amino acids of A. thaliana PhyB (PhyB 1-651 ) have been used as an optogenetic tool (Adrian et al, 2017; Baaske et al, 2019; Beyer et al, 2015; Beyer et al, 2018; Johnson and Toettcher, 2018; Levskaya et al, 2009; Müller et al, 2013b; Toettcher et al, 2013) and the photobiology of this fragment has been described previously (Smith et al, 2016). Here we used this PhyB form as a ligand.…”

Section: Resultsmentioning

confidence: 99%

“…With 740 nm light, PhyB undergoes a conformational transition to the OFF state preventing binding to PIF6. This light-dependent protein-protein interaction was utilized in several optogenetic applications (Kolar et al, 2018), such as the control of protein or organelle localization (Adrian et al, 2017; Beyer et al, 2018; Levskaya et al, 2009), intracellular signaling (Toettcher et al, 2013), nuclear transport of proteins (Beyer et al, 2015), cell adhesion (Baaske et al, 2019; Yüz et al, 2018) or gene expression (Müller et al, 2013a). Using high intensity light, the PhyB-PIF interaction can be switched ON and OFF within seconds (Levskaya et al, 2009; Mancinelli, 1994; Smith et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

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Optogenetic control shows that kinetic proofreading regulates the activity of the T cell receptor

Yousefi

Günther

Hörner

et al. 2019

eLife

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108

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The immune system distinguishes between self and foreign antigens. The kinetic proofreading (KPR) model proposes that T cells discriminate self from foreign ligands by the different ligand binding half-lives to the T cell receptor (TCR). It is challenging to test KPR as the available experimental systems fall short of only altering the binding half-lives and keeping other parameters of the interaction unchanged. We engineered an optogenetic system using the plant photoreceptor phytochrome B (PhyB) as a ligand to selectively control the dynamics of ligand binding to the TCR by light. This opto-ligand-TCR system was combined with the unique property of PhyB to continuously cycle between the binding and non-binding states under red light, with the light intensity determining the cycling rate and thus the binding duration. Mathematical modeling of our experimental datasets showed that indeed the ligand-TCR interaction half-life is the decisive factor for activating downstream TCR signaling, substantiating KPR.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Optogenetic control shows that kinetic proofreading regulates the activity of the T cell receptor

Yousefi

Günther

Hörner

et al. 2019

eLife

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108

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“…This unique feature of the opto-ligand-TCR system could enable researchers to locally or timely restrict ligand-receptor interaction. Fusing PIF to other receptors would allow to control ligand binding to those receptors as well, as we previously demonstrated for integrins (Baaske et al, 2019). Further, our system could be used to investigate the signaling events that happen after ligand dissociation, which have been mostly neglected due to the lack of appropriate methods.…”

Section: Controlmentioning

confidence: 94%

Optogenetic Tuning of Ligand Binding to The Human T cell Receptor Using The opto-ligand-TCR System

Yousefi¹,

Hörner²,

Wess³

et al. 2020

BIO-PROTOCOL

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T cells are one major cell type of the immune system that use their T cell antigen receptor (TCR) to bind and respond to foreign molecules derived from pathogens. The ligand-TCR interaction half-lives determine stimulation outcome. Until recently, scientists relied on mutating either the TCR or its ligands to investigate how varying TCR-ligand interaction durations impacted on T cell activation. Our newly created opto-ligand-TCR system allowed us to precisely and reversibly control ligand binding to the TCR by light illumination. This system uses phytochrome B (PhyB) tetramers as a light-regulated TCR ligand. PhyB can be photoconverted between a binding (ON) and non-binding (OFF) conformation by 660 nm and 740 nm light illumination, respectively. PhyB ON is able to bind to a synthetic TCR, generated by fusing the PhyB interacting factor (PIF) to the TCRβ chain. Switching PhyB to the OFF conformation disrupts this interaction. Sufficiently long binding of PhyB tetramers to the PIF-TCR led to T cell activation as measured by calcium influx. Here, we describe protocols for how to generate the tetrameric ligand for our opto-ligand-TCR system, how to measure ligand-TCR binding by flow cytometry and how to quantify T cell activation via calcium influx.

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“…Changes in cell contractility were found to be related to modifications in the transcriptional regulator YAP, demonstrating the ability of optogenetic approach to control mechanotransduction signaling pathways. More recently, Baaske and coworkers reported an optogenetic system based on an integrin engineered with a phytochrome-interacting factor domain (OptoIntegrin) and a red light-switchable phytochrome B-functionalized matrix (OptoMatrix) (Baaske et al, 2019). This receptor-ligand pair enables a reversible optogenetic control of integrin-matrix interaction, as well as the controlled activation of downstream mechanosensory signaling pathways.…”

Section: Optogenetic Methods To Interrogate Mechanotransduction Pathwaysmentioning

confidence: 99%

Innovative Tools for Mechanobiology: Unraveling Outside-In and Inside-Out Mechanotransduction

Mohammed

Versaevel

Bruyère

et al. 2019

Front. Bioeng. Biotechnol.

135

124

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Cells and tissues can sense and react to the modifications of the physico-chemical properties of the extracellular environment (ECM) through integrin-based adhesion sites and adapt their physiological response in a process called mechanotransduction. Due to their critical localization at the cell-ECM interface, transmembrane integrins are mediators of bidirectional signaling, playing a key role in “outside-in” and “inside-out” signal transduction. After presenting the basic conceptual fundamentals related to cell mechanobiology, we review the current state-of-the-art technologies that facilitate the understanding of mechanotransduction signaling pathways. Finally, we highlight innovative technological developments that can help to advance our understanding of the mechanisms underlying nuclear mechanotransduction.

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Optogenetic control of integrin-matrix interaction

Cited by 32 publications

References 38 publications

Optogenetic control shows that kinetic proofreading regulates the activity of the T cell receptor

Optogenetic control shows that kinetic proofreading regulates the activity of the T cell receptor

Optogenetic Tuning of Ligand Binding to The Human T cell Receptor Using The opto-ligand-TCR System

Innovative Tools for Mechanobiology: Unraveling Outside-In and Inside-Out Mechanotransduction

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