Active and inactive β1 integrins segregate into distinct nanoclusters in focal adhesions

Spiess, Matthias; Hernández-Varas, Pablo; Oddone, Anna; Olofsson, Helene; Blom, Hans; Waithe, Dominic; Lock, John G.; Lakadamyali, Melike; Strömblad, Staffan

doi:10.1083/jcb.201707075

Cited by 89 publications

(103 citation statements)

References 63 publications

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“…In particular, β5-integrin was found to assemble high-density ring-like clustering at the edges of adhesions while β1 or αV-integrins were found to be more homogeneously distributed. Within classical focal adhesion, distinct integrin heterodimers display different dynamics (Rossier et al, 2012) or cluster differently depending on their activation states (Spiess et al, 2018). However, the spatial segregation of specific integrin heterodimers appears to be a unique feature of cornerstone adhesions.…”

Section: Discussionmentioning

confidence: 99%

Superresolution architecture of pluripotency guarding adhesions

Stubb

Guzmán

Närvä

et al. 2018

Preprint

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Human pluripotent stem cells (hPSC) can generate almost all adult cell lineages. While it is clear that key transcriptional programmes are important elements for maintaining pluripotency, the equally essential requirement for cell adhesion to specific extracellular matrix components remains poorly defined. Our recent observation that hPSC colonies form unusually large "cornerstone" focal adhesions (FA), distinct from parental somatic cells, that are lost following differentiation, emphasises the potential of these atypical FA as gatekeepers of pluripotency. Here, using nanopatterns, we further demonstrate that physical restriction of adhesion size, in hPSC colonies, is sufficient to trigger differentiation. Using superresolution two-colour interferometric photoactivated localization microscopy (iPALM), we examined the three-dimensional architecture of these cornerstone adhesions and report vertical lamination of FA proteins with three main structural peculiarities: 1) integrin β5 and talin are present at high density, at the edges of cornerstone FA, adjacent to a vertical kank-rich protein wall. 2) Vinculin localises higher than expected with respect to the substrata, and 3) surprisingly, actin and α-actinin are present in two discrete layers, a previously undescribed localisation for these proteins. Finally, we report that depletion of kanks diminishes FA patterning, and actin organisation within the colony, indicating a key role for kanks in hPSC colony architecture.

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Section: Discussionmentioning

confidence: 99%

Superresolution architecture of pluripotency guarding adhesions

Stubb

Guzmán

Närvä

et al. 2018

Preprint

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“…Upregulating the Integrin b1/FAK/ERK Signaling and Subsequent Matrix Metalloproteinase (MMP)-9 Expression To further unravel the mechanism underlying PKM2-promoted cell invasiveness, we detected the activity of integrin b1 and its downstream signaling. The influence of secreted PKM2 on integrin b1 was investigated using the antibody 12G10, which specifically recognizes the active conformation of integrin b1 (Spiess et al, 2018). Obviously, r-PKM2 induced the activation of integrin b1, without affecting total integrin b1 level ( Figures 6A-6C).…”

Section: Secreted Pkm2 Exerts the Pro-metastasis Function Bymentioning

confidence: 99%

Secreted Pyruvate Kinase M2 Promotes Lung Cancer Metastasis through Activating the Integrin Beta1/FAK Signaling Pathway

et al. 2020

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Graphical Abstract Highlights d Secreted PKM2 is a potential serum biomarker for diagnosing lung cancer d Secreted PKM2 promotes lung cancer progression d Integrin b1 functions as a receptor for secreted PKM2 d Targeting secreted PKM2 and integrin b1 can attenuate lung cancer metastasis SUMMARYCancer cell-derived secretomes have been documented to play critical roles in cancer progression. Intriguingly, alternative extracellular roles of intracellular proteins are involved in various steps of tumor progression, which can offer strategies to fight cancer. Herein, we identify lung cancer progression-associated secretome signatures using mass spectrometry analysis. Among them, PKM2 is verified to be highly expressed and secreted in lung cancer cells and clinical samples. Functional analyses demonstrates that secreted PKM2 facilitates tumor metastasis. Furthermore, mass spectrometry analysis and functional validation identify integrin b1 as a receptor of secreted PKM2. Mechanistically, secreted PKM2 directly bound to integrin b1 and subsequently activated the FAK/SRC/ERK axis to promote tumor metastasis. Collectively, our findings suggest that PKM2 is a potential serum biomarker for diagnosing lung cancer and that targeting the secreted PKM2-integrin b1 axis can inhibit lung cancer development, which provides evidence of a potential therapeutic strategy in lung cancer.

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“…To determine if the GIV•K2 interaction persists later in mature FAs, we used two-color super-resolution stimulated emission depletion (STED) microscopy (Hell and Wichmann, 1994;Klar et al, 2000) enabling a lateral resolution of ~40 nm to assess nanoscale colocalization of endogenous pYGIV and K2 within focal adhesions. Prior studies using STED (Colin-York et al, 2017;Spiess et al, 2018) have revealed the superiority of this approach over conventional microscopy for assessing the organization of FAs into nanometer size clusters of multi-protein assemblies. We observed nanoscale co-localization between pYGIV and K2 within mature FA-structures that were positive for paxillin [ Fig 5B]; compared to GIV-WT cells, such structures were far fewer and virtually lacking in cells expressing the PGxA mutant.…”

Section: The Giv•kindlin-2 Interaction Enhances Tumor Cell Adhesion mentioning

confidence: 99%

GIV•Kindlin interaction is required for Kindlin-Mediated Integrin Recognition and Activation

Rohena

Kalogriopoulos

Rajapakse

et al. 2019

Preprint

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The eTOC blurb (50 words)Integrins mediate cell adhesion to the extracellular matrix; their dysregulation fuels inflammation, cancer cell invasion and metastasis. Authors show how two pro-metastatic scaffold proteins, Kindlin and GIV/Girdin bind and cooperatively enhance their allosteric coupling to integrins, and their subsequent activation.Findings reveal novel interfaces in integrin signaling for pharmacologic manipulation. HIGHLIGHTS• GIV and Kindlin(K2), two integrin adaptors that promote metastasis, bind each other • Binding of GIV or integrin to K2 allosterically enhances GIV•K2•integrin complexes• Binding is required for the maximal recruitment of GIV and K2 to active integrins • Binding facilitates integrin clustering, activation, tumor cell adhesion, invasion. ABSTRACT (150 words)Cells perceive and respond to the extracellular matrix (ECM) via integrin receptors; their dysregulation has been implicated in inflammation and cancer metastasis. Here we show that a guanine nucleotide exchange modulator of trimeric-GTPase Gαi, GIV (a.k.a Girdin), directly binds the integrin adaptor Kindlin-2. A noncanonical short linear motif within GIV's C-terminus binds Kindlin-2-FERM3 domain at a site that is distinct from the binding site for the canonical NPxY motif on the -integrin tail. Binding of GIV to Kindlin-2 allosterically enhances Kindlin-2's affinity for β 1-integrin. Consequently, integrin activation and clustering are maximized, which augments cell adhesion, spreading and invasion. Findings elucidate how the GIV•Kindlin-2 complex has a two-fold impact: it allosterically synergizes integrin activation and enables β 1-integrins to indirectly access and modulate trimeric GTPases via the complex. Furthermore, Cox proportional-hazard models on tumor transcriptomics provide trans-scale evidence of synergistic interactions between GIV•Kindlin-2•β1-integrin on time to progression to metastasis.

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Active and inactive β1 integrins segregate into distinct nanoclusters in focal adhesions

Cited by 89 publications

References 63 publications

Superresolution architecture of pluripotency guarding adhesions

Superresolution architecture of pluripotency guarding adhesions

Secreted Pyruvate Kinase M2 Promotes Lung Cancer Metastasis through Activating the Integrin Beta1/FAK Signaling Pathway

GIV•Kindlin interaction is required for Kindlin-Mediated Integrin Recognition and Activation

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