LFA-1 activates focal adhesion kinases FAK1/PYK2 to generate LAT-GRB2-SKAP1 complexes that terminate T-cell conjugate formation

Raab, Monika; Lu, Yuning; Köhler, Karsten; Smith, Xin; Strebhardt, Klaus; Rudd, Christopher E.

doi:10.1038/ncomms16001

Cited by 32 publications

(52 citation statements)

References 73 publications

(141 reference statements)

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“…4D). Additionally, FAK has established roles in T-cell signaling (41,42). Previous reports demonstrated that PI(4,5)P2 binding activated FAK activity (35,43,44).…”

Section: Weak Tcr Signals Activate Fak Via Elevated Pi(45)p2 In Cd4 mentioning

confidence: 99%

T cells transduce T-cell receptor signal strength by generating different phosphatidylinositols

Hawse

Cattley

2019

Journal of Biological Chemistry

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Edited by Peter Cresswell T-cell receptor (TCR) signaling strength is a dominant factor regulating T-cell differentiation, thymic development, and cytokine signaling. The molecular mechanisms by which TCR signal strength is transduced to downstream signaling networks remains ill-defined. Using computational modeling, biochemical assays, and imaging flow cytometry, we found here that TCR signal strength differentially generates phosphatidylinositol species. Weak TCR signals generated elevated phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) and reduced phosphatidylinositol (3,4,5)-trisphosphate (PIP3) levels, whereas strong TCR signals reduced PI(4,5)P2 and elevated PIP3 levels. A proteomics screen revealed that focal adhesion kinase bound PI(4,5)P2, biochemical assays disclosed that focal adhesion kinase is preferentially activated by weak TCR signals and is required for optimal Treg induction, and further biochemical experiments revealed how TCR signaling strength regulates AKT activation. Low PIP3 levels generated by weak TCR signals were sufficient to activate phosphoinositide-dependent kinase-1 to phosphorylate AKT on Thr-308 but insufficient to activate mTOR complex 2 (mTORC2), whereas elevated PIP3 levels generated by a strong TCR signal were required to activate mTORC2 to phosphorylate Ser-473 on AKT. Our results provide support for a model that links TCR signaling to mTORC2 activation via phosphoinositide 3-kinase signaling. Together, the findings in this work establish that T cells measure TCR signal strength by generating different levels of phosphatidylinositol species that engage alternate signaling networks to control cell fate decisions. cro ARTICLE

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“…4D). Additionally, FAK has established roles in T-cell signaling (41,42). Previous reports demonstrated that PI(4,5)P2 binding activated FAK activity (35,43,44).…”

Section: Weak Tcr Signals Activate Fak Via Elevated Pi(45)p2 In Cd4 mentioning

confidence: 99%

T cells transduce T-cell receptor signal strength by generating different phosphatidylinositols

Hawse

Cattley

2019

Journal of Biological Chemistry

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“…FAK is basally activated by the Src family kinase LCK in effector T-cells at Y576 or Y577 (depending on species), and Y925. This primes FAK for full activation though phosphorylation at the autophosphorylation site Y397 (herein denoted pY397 FAK) (Chapman and Houtman, 2014), which in turn regulates LFA-1 adhesion and de-adhesion (Raab et al, 2017). Expression of dominant-negative mutants of FAK in T-cells reduces the speed of migration on ICAM-1-coated glass (Rose et al, 2003), and overexpression of a negative regulator of FAK phosphorylation impairs LFA-1 clustering and reduces ICAM-1-LFA-1-driven T-cell homo-aggregation (Giannoni et al, 2003).…”

Section: The Speed Of Actin Retrograde Flow Is Positively Correlated mentioning

confidence: 99%

“…Control of adhesion is achieved through actin linkers (talin and vinculin) and regulatory kinases [FAK (also known as PTK2) and Src family kinases] (Nordenfelt et al, 2016;Raab et al, 2017), a mechanism originally described in other, much more slowly migrating cell types that characteristically form large micron sized focal adhesions (Kanchanawong et al, 2010). These components were originally described in slow migrating adherent cells that characteristically form large micron-sized focal adhesions (Kanchanawong et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Differential nanoscale organisation of LFA-1 modulates T-cell migration

Shannon

Pineau

Griffié

et al. 2019

Journal of Cell Science

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Effector T-cells rely on integrins to drive adhesion and migration to facilitate their immune function. The heterodimeric transmembrane integrin LFA-1 (αLβ2 integrin) regulates adhesion and migration of effector T-cells through linkage of the extracellular matrix with the intracellular actin treadmill machinery. Here, we quantified the velocity and direction of F-actin flow in migrating T-cells alongside singlemolecule localisation of transmembrane and intracellular LFA-1. Results showed that actin retrograde flow positively correlated and immobile actin negatively correlated with T-cell velocity. Plasma membrane-localised LFA-1 forms unique nano-clustering patterns in the leading edge, compared to the mid-focal zone, of migrating Tcells. Deleting the cytosolic phosphatase PTPN22, loss-of-function mutations of which have been linked to autoimmune disease, increased T-cell velocity, and leading-edge co-clustering of pY397 FAK, pY416 Src family kinases and LFA-1. These data suggest that differential nanoclustering patterns of LFA-1 in migrating T-cells may instruct intracellular signalling. Our data presents a paradigm where T-cells modulate the nanoscale organisation of adhesion and signalling molecules to fine tune their migration speed, with implications for the regulation of immune and inflammatory responses. This article has an associated First Person interview with the first author of the paper.

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“…FAK is basally activated by Src-family kinase LCK in effector T-cells at Y576/577 and Y925. This primes FAK for full activation at the auto-phosphorylation site pY397 (Chapman & Houtman 2014) which in turn regulates LFA-1 adhesion and deadhesion (Raab et al 2017b). Expression of dominant-negative mutants of FAK in T-cells reduce the speed of migration on ICAM-1 coated glass (Rose et al 2003), and overexpression of a negative regulator of FAK phosphorylation impairs LFA-1 clustering and reduces ICAM-1/LFA-1 driven Tcell homo-aggregation (Giannoni et al 2003).…”

Section: Deleting Ptpn22 Increases T-cell Velocity the Size Of 3d Pymentioning

confidence: 99%

“…Control of adhesion is also achieved through actin linkers (talin, vinculin) and regulatory kinases (FAK, Src family kinases) (Nordenfelt et al 2016;Raab et al 2017a), common to other migrating cell types, that form large micron sized focal adhesions (Kanchanawong et al 2010). In T cells, these complexes are much smaller (nanoscale), containing only a few molecules (Burn et al 2016) but similar in size and composition to nascent adhesions (Changede et al 2015).…”

Section: Introductionmentioning

confidence: 99%

Differential nanoscale organisation of LFA-1 modulates T cell migration

Shannon

Pineau

Griffié

et al. 2019

Preprint

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Effector T-cells rely on integrins to drive adhesion and migration to facilitate their immune function. Heterodimeric transmembrane integrin LFA-1 (αLβ2) regulates adhesion and migration through linkage of the extracellular matrix with the intracellular actin treadmill machinery. We quantitated the velocity and direction of F-actin flow in migrating T-cells alongside single molecule localisation of transmembrane and intracellular LFA-1. Our results show that retrograde actin flow positively correlated and immobile actin negatively correlated with T-cell velocity. Plasma membrane localised LFA-1 forms unique nano-clustering patterns in the leading edge, compared to the mid-focal zone, in migrating T-cells. Deleting the cytosolic phosphatase PTPN22, a negative regulator of integrin signaling, increased T-cell velocity, and leading-edge cluster co-localisation of pY397 FAK, pY416 Src family kinases and LFA-1. These data suggest that differential nanoclustering patterns of LFA-1 in migrating T-cells can instruct intracellular signalling linked with the actin treadmill. Our data presents a paradigm where T cells modulate the nanoscale organisation of adhesion and signalling molecules to fine tune their migration speed. This has implications for the regulation of immune and inflammatory responses. ResultsThe speed of actin retrograde flow is positively correlated with T-cell velocity and negatively correlated with immobile actin.

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LFA-1 activates focal adhesion kinases FAK1/PYK2 to generate LAT-GRB2-SKAP1 complexes that terminate T-cell conjugate formation

Cited by 32 publications

References 73 publications

T cells transduce T-cell receptor signal strength by generating different phosphatidylinositols

T cells transduce T-cell receptor signal strength by generating different phosphatidylinositols

Differential nanoscale organisation of LFA-1 modulates T-cell migration

Differential nanoscale organisation of LFA-1 modulates T cell migration

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