Phosphatidylinositol 4,5-bisphosphate triggers activation of focal adhesion kinase by inducing clustering and conformational changes

Guillermina M Goñi; Carolina Epifano; Jasminka Boskovic; Marta Camacho-Artacho; Jing Zhou; Agnieszka Bronowska; M Teresa Martín; Michael J Eck; Leonor Kremer; Frauke Gräter; Francesco Luigi Gervasio; Mirna Perez-Moreno; Daniel Lietha

doi:10.1073/pnas.1317022111

Phosphatidylinositol 4,5-bisphosphate triggers activation of focal adhesion kinase by inducing clustering and conformational changes

Guillermina M Goñi, Carolina Epifano, Jasminka Boskovic, Marta Camacho-Artacho, Jing Zhou, Agnieszka Bronowska, M Teresa Martín, Michael J Eck, Leonor Kremer, Frauke Gräter, Francesco Luigi Gervasio, Mirna Perez-Moreno, Daniel Lietha

70 Citationer (Scopus)

Abstract

Focal adhesion kinase (FAK) is a nonreceptor tyrosine kinase (NRTK) with key roles in integrating growth and cell matrix adhesion signals, and FAK is a major driver of invasion and metastasis in cancer. Cell adhesion via integrin receptors is well known to trigger FAK signaling, and many of the players involved are known; however, mechanistically, FAK activation is not understood. Here, using a multidisciplinary approach, including biochemical, biophysical, structural, computational, and cell biology approaches, we provide a detailed view of a multistep activation mechanism of FAK initiated by phosphatidylinositol-4,5-bisphosphate [PI(4,5)P2]. Interestingly, the mechanism differs from canonical NRTK activation and is tailored to the dual catalytic and scaffolding function of FAK. We find PI(4,5)P2 induces clustering of FAK on the lipid bilayer by binding a basic region in the regulatory 4.1, ezrin, radixin, moesin homology (FERM) domain. In these clusters, PI(4,5)P2 induces a partially open FAK conformation where the autophosphorylation site is exposed, facilitating efficient autophosphorylation and subsequent Src recruitment. However, PI(4,5)P2 does not release autoinhibitory interactions; rather, Src phosphorylation of the activation loop in FAK results in release of the FERM/kinase tether and full catalytic activation. We propose that PI(4,5)P2 and its generation in focal adhesions by the enzyme phosphatidylinositol 4-phosphate 5-kinase type Iγ are important in linking integrin signaling to FAK activation.

Originalsprog	Engelsk
Tidsskrift	Proceedings of the National Academy of Sciences of the United States of America
Vol/bind	111
Udgave nummer	31
Sider (fra-til)	E3177-E3186
ISSN	0027-8424
DOI	https://doi.org/10.1073/pnas.1317022111
Status	Udgivet - 5 aug. 2014
Udgivet eksternt	Ja

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Goñi, G. M., Epifano, C., Boskovic, J., Camacho-Artacho, M., Zhou, J., Bronowska, A., Martín, M. T., Eck, M. J., Kremer, L., Gräter, F., Gervasio, F. L., Perez-Moreno, M., & Lietha, D. (2014). Phosphatidylinositol 4,5-bisphosphate triggers activation of focal adhesion kinase by inducing clustering and conformational changes. Proceedings of the National Academy of Sciences of the United States of America, 111(31), E3177-E3186. https://doi.org/10.1073/pnas.1317022111

Phosphatidylinositol 4,5-bisphosphate triggers activation of focal adhesion kinase by inducing clustering and conformational changes. / Goñi, Guillermina M; Epifano, Carolina; Boskovic, Jasminka et al.

I: Proceedings of the National Academy of Sciences of the United States of America, Bind 111, Nr. 31, 05.08.2014, s. E3177-E3186.

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

Goñi, GM, Epifano, C, Boskovic, J, Camacho-Artacho, M, Zhou, J, Bronowska, A, Martín, MT, Eck, MJ, Kremer, L, Gräter, F, Gervasio, FL, Perez-Moreno, M & Lietha, D 2014, 'Phosphatidylinositol 4,5-bisphosphate triggers activation of focal adhesion kinase by inducing clustering and conformational changes', Proceedings of the National Academy of Sciences of the United States of America, bind 111, nr. 31, s. E3177-E3186. https://doi.org/10.1073/pnas.1317022111

@article{e015a8fc7d65461595b4511bdda0ea0b,

title = "Phosphatidylinositol 4,5-bisphosphate triggers activation of focal adhesion kinase by inducing clustering and conformational changes",

abstract = "Focal adhesion kinase (FAK) is a nonreceptor tyrosine kinase (NRTK) with key roles in integrating growth and cell matrix adhesion signals, and FAK is a major driver of invasion and metastasis in cancer. Cell adhesion via integrin receptors is well known to trigger FAK signaling, and many of the players involved are known; however, mechanistically, FAK activation is not understood. Here, using a multidisciplinary approach, including biochemical, biophysical, structural, computational, and cell biology approaches, we provide a detailed view of a multistep activation mechanism of FAK initiated by phosphatidylinositol-4,5-bisphosphate [PI(4,5)P2]. Interestingly, the mechanism differs from canonical NRTK activation and is tailored to the dual catalytic and scaffolding function of FAK. We find PI(4,5)P2 induces clustering of FAK on the lipid bilayer by binding a basic region in the regulatory 4.1, ezrin, radixin, moesin homology (FERM) domain. In these clusters, PI(4,5)P2 induces a partially open FAK conformation where the autophosphorylation site is exposed, facilitating efficient autophosphorylation and subsequent Src recruitment. However, PI(4,5)P2 does not release autoinhibitory interactions; rather, Src phosphorylation of the activation loop in FAK results in release of the FERM/kinase tether and full catalytic activation. We propose that PI(4,5)P2 and its generation in focal adhesions by the enzyme phosphatidylinositol 4-phosphate 5-kinase type Iγ are important in linking integrin signaling to FAK activation.",

keywords = "Adenosine Triphosphate, Allosteric Regulation, Amino Acid Sequence, Amino Acids, Biocatalysis, Cell Adhesion, Cluster Analysis, Enzyme Activation, Fluorescence Resonance Energy Transfer, Focal Adhesion Protein-Tyrosine Kinases, Gene Knockdown Techniques, HeLa Cells, Humans, Models, Molecular, Molecular Sequence Data, Mutant Proteins, Phosphatidylinositol 4,5-Diphosphate, Phosphorylation, Phosphotransferases (Alcohol Group Acceptor), Protein Binding, Protein Structure, Tertiary, Signal Transduction, src-Family Kinases, Journal Article, Research Support, Non-U.S. Gov't",

author = "Go{\~n}i, {Guillermina M} and Carolina Epifano and Jasminka Boskovic and Marta Camacho-Artacho and Jing Zhou and Agnieszka Bronowska and Mart{\'i}n, {M Teresa} and Eck, {Michael J} and Leonor Kremer and Frauke Gr{\"a}ter and Gervasio, {Francesco Luigi} and Mirna Perez-Moreno and Daniel Lietha",

year = "2014",

month = aug,

day = "5",

doi = "10.1073/pnas.1317022111",

language = "English",

volume = "111",

pages = "E3177--E3186",

journal = "Proceedings of the National Academy of Sciences of the United States of America",

issn = "0027-8424",

publisher = "The National Academy of Sciences of the United States of America",

number = "31",

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TY - JOUR

T1 - Phosphatidylinositol 4,5-bisphosphate triggers activation of focal adhesion kinase by inducing clustering and conformational changes

AU - Goñi, Guillermina M

AU - Epifano, Carolina

AU - Boskovic, Jasminka

AU - Camacho-Artacho, Marta

AU - Zhou, Jing

AU - Bronowska, Agnieszka

AU - Martín, M Teresa

AU - Eck, Michael J

AU - Kremer, Leonor

AU - Gräter, Frauke

AU - Gervasio, Francesco Luigi

AU - Perez-Moreno, Mirna

AU - Lietha, Daniel

PY - 2014/8/5

Y1 - 2014/8/5

N2 - Focal adhesion kinase (FAK) is a nonreceptor tyrosine kinase (NRTK) with key roles in integrating growth and cell matrix adhesion signals, and FAK is a major driver of invasion and metastasis in cancer. Cell adhesion via integrin receptors is well known to trigger FAK signaling, and many of the players involved are known; however, mechanistically, FAK activation is not understood. Here, using a multidisciplinary approach, including biochemical, biophysical, structural, computational, and cell biology approaches, we provide a detailed view of a multistep activation mechanism of FAK initiated by phosphatidylinositol-4,5-bisphosphate [PI(4,5)P2]. Interestingly, the mechanism differs from canonical NRTK activation and is tailored to the dual catalytic and scaffolding function of FAK. We find PI(4,5)P2 induces clustering of FAK on the lipid bilayer by binding a basic region in the regulatory 4.1, ezrin, radixin, moesin homology (FERM) domain. In these clusters, PI(4,5)P2 induces a partially open FAK conformation where the autophosphorylation site is exposed, facilitating efficient autophosphorylation and subsequent Src recruitment. However, PI(4,5)P2 does not release autoinhibitory interactions; rather, Src phosphorylation of the activation loop in FAK results in release of the FERM/kinase tether and full catalytic activation. We propose that PI(4,5)P2 and its generation in focal adhesions by the enzyme phosphatidylinositol 4-phosphate 5-kinase type Iγ are important in linking integrin signaling to FAK activation.

AB - Focal adhesion kinase (FAK) is a nonreceptor tyrosine kinase (NRTK) with key roles in integrating growth and cell matrix adhesion signals, and FAK is a major driver of invasion and metastasis in cancer. Cell adhesion via integrin receptors is well known to trigger FAK signaling, and many of the players involved are known; however, mechanistically, FAK activation is not understood. Here, using a multidisciplinary approach, including biochemical, biophysical, structural, computational, and cell biology approaches, we provide a detailed view of a multistep activation mechanism of FAK initiated by phosphatidylinositol-4,5-bisphosphate [PI(4,5)P2]. Interestingly, the mechanism differs from canonical NRTK activation and is tailored to the dual catalytic and scaffolding function of FAK. We find PI(4,5)P2 induces clustering of FAK on the lipid bilayer by binding a basic region in the regulatory 4.1, ezrin, radixin, moesin homology (FERM) domain. In these clusters, PI(4,5)P2 induces a partially open FAK conformation where the autophosphorylation site is exposed, facilitating efficient autophosphorylation and subsequent Src recruitment. However, PI(4,5)P2 does not release autoinhibitory interactions; rather, Src phosphorylation of the activation loop in FAK results in release of the FERM/kinase tether and full catalytic activation. We propose that PI(4,5)P2 and its generation in focal adhesions by the enzyme phosphatidylinositol 4-phosphate 5-kinase type Iγ are important in linking integrin signaling to FAK activation.

KW - Adenosine Triphosphate

KW - Allosteric Regulation

KW - Amino Acid Sequence

KW - Amino Acids

KW - Biocatalysis

KW - Cell Adhesion

KW - Cluster Analysis

KW - Enzyme Activation

KW - Fluorescence Resonance Energy Transfer

KW - Focal Adhesion Protein-Tyrosine Kinases

KW - Gene Knockdown Techniques

KW - HeLa Cells

KW - Humans

KW - Models, Molecular

KW - Molecular Sequence Data

KW - Mutant Proteins

KW - Phosphatidylinositol 4,5-Diphosphate

KW - Phosphorylation

KW - Phosphotransferases (Alcohol Group Acceptor)

KW - Protein Binding

KW - Protein Structure, Tertiary

KW - Signal Transduction

KW - src-Family Kinases

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1073/pnas.1317022111

DO - 10.1073/pnas.1317022111

M3 - Journal article

C2 - 25049397

SN - 0027-8424

VL - 111

SP - E3177-E3186

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 31

ER -

Phosphatidylinositol 4,5-bisphosphate triggers activation of focal adhesion kinase by inducing clustering and conformational changes

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