Specific interactions of neuronal focal adhesion kinase isoforms with Src kinases and amphiphysin

Samantha Messina, Franco Onofri, Lucilla Bongiorno-Borbone, Silvia Giovedì, Flavia Valtorta, Jean Antoine Girault, Fabio Benfenati

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Focal adhesion kinase (FAK) is a non-receptor tyrosine kinase that activates Src family kinases via SH2- and SH3-mediated interactions. Specific FAK isoforms (FAK+), responsive to depolarization and neurotransmitters, are enriched in neurons. We analyzed the interactions of endogenous FAK+ and recombinant FAK+ isoforms containing amino acid insertions (boxes 6,7,28) with an array of SH3 domains and the c-Src SH2/SH3 domain tandem. Endogenous FAK+ bound specifically to the SH3 domains of c-Src (but not n-Src), Fyn, Yes, phosphtidylinositol-3 kinase, amphiphysin II, amphiphysin I, phospholipase Cγ and NH2-terminal Grb2. The inclusion of boxes 6,7 was associated with a significant decrease in the binding of FAK+ to the c-Src and Fyn SH3 domains, and a significant increase in the binding to the Src SH2 domain, as a consequence of the higher phosphorylation of Tyr-397. The novel interaction with the amphiphysin SH3 domain, involving the COOH-terminal proline-rich region of FAK, was confirmed by coimmunoprecipitation of the two proteins and a closely similar response to stimuli affecting the actin cytoskeleton. Moreover, an impairment of endocytosis was observed in synaptosomes after internalization of a proline-rich peptide corresponding to the site of interaction. The data account for the different subcellular distribution of FAK and Src kinases and the specific regulation of the transduction pathways linked to FAK activation in the brain and implicate FAK in the regulation of membrane trafficking in nerve terminals.

Original languageEnglish
Pages (from-to)253-265
Number of pages13
JournalJournal of Neurochemistry
Volume84
Issue number2
DOIs
Publication statusPublished - Jan 2003

Fingerprint

Focal Adhesion Protein-Tyrosine Kinases
src-Family Kinases
Protein Isoforms
src Homology Domains
Proline
amphiphysin
Phosphorylation
Synaptosomes
Depolarization
Type C Phospholipases
Endocytosis
Actin Cytoskeleton
Protein-Tyrosine Kinases
Neurons
Neurotransmitter Agents
Actins
Brain
Phosphotransferases
Chemical activation

Keywords

  • Amphiphysin
  • Nerve terminals
  • SH2 domains
  • SH3 domains
  • Tyrosine phosphorylation

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Specific interactions of neuronal focal adhesion kinase isoforms with Src kinases and amphiphysin. / Messina, Samantha; Onofri, Franco; Bongiorno-Borbone, Lucilla; Giovedì, Silvia; Valtorta, Flavia; Girault, Jean Antoine; Benfenati, Fabio.

In: Journal of Neurochemistry, Vol. 84, No. 2, 01.2003, p. 253-265.

Research output: Contribution to journalArticle

Messina, S, Onofri, F, Bongiorno-Borbone, L, Giovedì, S, Valtorta, F, Girault, JA & Benfenati, F 2003, 'Specific interactions of neuronal focal adhesion kinase isoforms with Src kinases and amphiphysin', Journal of Neurochemistry, vol. 84, no. 2, pp. 253-265. https://doi.org/10.1046/j.1471-4159.2003.01519.x
Messina, Samantha ; Onofri, Franco ; Bongiorno-Borbone, Lucilla ; Giovedì, Silvia ; Valtorta, Flavia ; Girault, Jean Antoine ; Benfenati, Fabio. / Specific interactions of neuronal focal adhesion kinase isoforms with Src kinases and amphiphysin. In: Journal of Neurochemistry. 2003 ; Vol. 84, No. 2. pp. 253-265.
@article{8270b515cd5a4fb190533722f5ab39d7,
title = "Specific interactions of neuronal focal adhesion kinase isoforms with Src kinases and amphiphysin",
abstract = "Focal adhesion kinase (FAK) is a non-receptor tyrosine kinase that activates Src family kinases via SH2- and SH3-mediated interactions. Specific FAK isoforms (FAK+), responsive to depolarization and neurotransmitters, are enriched in neurons. We analyzed the interactions of endogenous FAK+ and recombinant FAK+ isoforms containing amino acid insertions (boxes 6,7,28) with an array of SH3 domains and the c-Src SH2/SH3 domain tandem. Endogenous FAK+ bound specifically to the SH3 domains of c-Src (but not n-Src), Fyn, Yes, phosphtidylinositol-3 kinase, amphiphysin II, amphiphysin I, phospholipase Cγ and NH2-terminal Grb2. The inclusion of boxes 6,7 was associated with a significant decrease in the binding of FAK+ to the c-Src and Fyn SH3 domains, and a significant increase in the binding to the Src SH2 domain, as a consequence of the higher phosphorylation of Tyr-397. The novel interaction with the amphiphysin SH3 domain, involving the COOH-terminal proline-rich region of FAK, was confirmed by coimmunoprecipitation of the two proteins and a closely similar response to stimuli affecting the actin cytoskeleton. Moreover, an impairment of endocytosis was observed in synaptosomes after internalization of a proline-rich peptide corresponding to the site of interaction. The data account for the different subcellular distribution of FAK and Src kinases and the specific regulation of the transduction pathways linked to FAK activation in the brain and implicate FAK in the regulation of membrane trafficking in nerve terminals.",
keywords = "Amphiphysin, Nerve terminals, SH2 domains, SH3 domains, Tyrosine phosphorylation",
author = "Samantha Messina and Franco Onofri and Lucilla Bongiorno-Borbone and Silvia Gioved{\`i} and Flavia Valtorta and Girault, {Jean Antoine} and Fabio Benfenati",
year = "2003",
month = "1",
doi = "10.1046/j.1471-4159.2003.01519.x",
language = "English",
volume = "84",
pages = "253--265",
journal = "Journal of Neurochemistry",
issn = "0022-3042",
publisher = "Wiley-Blackwell",
number = "2",

}

TY - JOUR

T1 - Specific interactions of neuronal focal adhesion kinase isoforms with Src kinases and amphiphysin

AU - Messina, Samantha

AU - Onofri, Franco

AU - Bongiorno-Borbone, Lucilla

AU - Giovedì, Silvia

AU - Valtorta, Flavia

AU - Girault, Jean Antoine

AU - Benfenati, Fabio

PY - 2003/1

Y1 - 2003/1

N2 - Focal adhesion kinase (FAK) is a non-receptor tyrosine kinase that activates Src family kinases via SH2- and SH3-mediated interactions. Specific FAK isoforms (FAK+), responsive to depolarization and neurotransmitters, are enriched in neurons. We analyzed the interactions of endogenous FAK+ and recombinant FAK+ isoforms containing amino acid insertions (boxes 6,7,28) with an array of SH3 domains and the c-Src SH2/SH3 domain tandem. Endogenous FAK+ bound specifically to the SH3 domains of c-Src (but not n-Src), Fyn, Yes, phosphtidylinositol-3 kinase, amphiphysin II, amphiphysin I, phospholipase Cγ and NH2-terminal Grb2. The inclusion of boxes 6,7 was associated with a significant decrease in the binding of FAK+ to the c-Src and Fyn SH3 domains, and a significant increase in the binding to the Src SH2 domain, as a consequence of the higher phosphorylation of Tyr-397. The novel interaction with the amphiphysin SH3 domain, involving the COOH-terminal proline-rich region of FAK, was confirmed by coimmunoprecipitation of the two proteins and a closely similar response to stimuli affecting the actin cytoskeleton. Moreover, an impairment of endocytosis was observed in synaptosomes after internalization of a proline-rich peptide corresponding to the site of interaction. The data account for the different subcellular distribution of FAK and Src kinases and the specific regulation of the transduction pathways linked to FAK activation in the brain and implicate FAK in the regulation of membrane trafficking in nerve terminals.

AB - Focal adhesion kinase (FAK) is a non-receptor tyrosine kinase that activates Src family kinases via SH2- and SH3-mediated interactions. Specific FAK isoforms (FAK+), responsive to depolarization and neurotransmitters, are enriched in neurons. We analyzed the interactions of endogenous FAK+ and recombinant FAK+ isoforms containing amino acid insertions (boxes 6,7,28) with an array of SH3 domains and the c-Src SH2/SH3 domain tandem. Endogenous FAK+ bound specifically to the SH3 domains of c-Src (but not n-Src), Fyn, Yes, phosphtidylinositol-3 kinase, amphiphysin II, amphiphysin I, phospholipase Cγ and NH2-terminal Grb2. The inclusion of boxes 6,7 was associated with a significant decrease in the binding of FAK+ to the c-Src and Fyn SH3 domains, and a significant increase in the binding to the Src SH2 domain, as a consequence of the higher phosphorylation of Tyr-397. The novel interaction with the amphiphysin SH3 domain, involving the COOH-terminal proline-rich region of FAK, was confirmed by coimmunoprecipitation of the two proteins and a closely similar response to stimuli affecting the actin cytoskeleton. Moreover, an impairment of endocytosis was observed in synaptosomes after internalization of a proline-rich peptide corresponding to the site of interaction. The data account for the different subcellular distribution of FAK and Src kinases and the specific regulation of the transduction pathways linked to FAK activation in the brain and implicate FAK in the regulation of membrane trafficking in nerve terminals.

KW - Amphiphysin

KW - Nerve terminals

KW - SH2 domains

KW - SH3 domains

KW - Tyrosine phosphorylation

UR - http://www.scopus.com/inward/record.url?scp=0037265829&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037265829&partnerID=8YFLogxK

U2 - 10.1046/j.1471-4159.2003.01519.x

DO - 10.1046/j.1471-4159.2003.01519.x

M3 - Article

VL - 84

SP - 253

EP - 265

JO - Journal of Neurochemistry

JF - Journal of Neurochemistry

SN - 0022-3042

IS - 2

ER -