Myristoylation-Dependent Palmitoylation of the Receptor Tyrosine Kinase Adaptor FRS2α

Barbara Barylko, Yu Ju Chen, Jared Hennen, Isaac Angert, Yan Chen, Joachim D. Mueller, Hui Qiao Sun, Clinton A. Taylor, Jen Liou, Helen Yin, Joseph P. Albanesi

Research output: Contribution to journalArticle

Abstract

An early step in signaling from activated receptor tyrosine kinases (RTKs) is the recruitment of cytosolic adaptor proteins to autophosphorylated tyrosines in the receptor cytoplasmic domains. Fibroblast growth factor receptor substrate 2α (FRS2α) associates via its phosphotyrosine-binding domain (PTB) to FGF receptors (FGFRs). Upon FGFR activation, FRS2α undergoes phosphorylation on multiple tyrosines, triggering recruitment of the adaptor Grb2 and the tyrosine phosphatase Shp2, resulting in stimulation of PI3K/AKT and MAPK signaling pathways. FRS2α also undergoes N-myristoylation, which was shown to be important for its localization to membranes and its ability to stimulate downstream signaling events (Kouhara et al., 1997). Here we show that FRS2α is also palmitoylated in cells and that cysteines 4 and 5 account for the entire modification. We further show that mutation of those two cysteines interferes with FRS2α localization to the plasma membrane (PM), and we quantify this observation using fluorescence fluctuation spectroscopy approaches. Importantly, prevention of myristoylation by introduction of a G2A mutation also abrogates palmitoylation, raising the possibility that signaling defects previously ascribed to the G2A mutant may actually be due to a failure of that mutant to undergo palmitoylation. Our results demonstrate that FRS2α undergoes coupled myristoylation and palmitoylation. Unlike stable cotranslational modifications, such as myristoylation and prenylation, palmitoylation is reversible due to the relative lability of the thioester linkage. Therefore, palmitoylation may provide a mechanism, in addition to phosphorylation, for dynamic regulation of FRS2 and its downstream signaling pathways.

Original languageEnglish (US)
Pages (from-to)2809-2813
Number of pages5
JournalBiochemistry
Volume58
Issue number25
DOIs
StatePublished - Jun 25 2019

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Lipoylation
Receptor Protein-Tyrosine Kinases
Fibroblast Growth Factor Receptors
Substrates
Tyrosine
Cysteine
Phosphorylation
Receptor, Fibroblast Growth Factor, Type 2
Prenylation
Mutation
Phosphotyrosine
Fluorescence Spectrometry
Phosphatidylinositol 3-Kinases
Phosphoric Monoester Hydrolases
Cell membranes
Cytoplasmic and Nuclear Receptors
Cell Membrane
Membranes
Fluorescence
Chemical activation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Myristoylation-Dependent Palmitoylation of the Receptor Tyrosine Kinase Adaptor FRS2α. / Barylko, Barbara; Chen, Yu Ju; Hennen, Jared; Angert, Isaac; Chen, Yan; Mueller, Joachim D.; Sun, Hui Qiao; Taylor, Clinton A.; Liou, Jen; Yin, Helen; Albanesi, Joseph P.

In: Biochemistry, Vol. 58, No. 25, 25.06.2019, p. 2809-2813.

Research output: Contribution to journalArticle

Barylko, Barbara ; Chen, Yu Ju ; Hennen, Jared ; Angert, Isaac ; Chen, Yan ; Mueller, Joachim D. ; Sun, Hui Qiao ; Taylor, Clinton A. ; Liou, Jen ; Yin, Helen ; Albanesi, Joseph P. / Myristoylation-Dependent Palmitoylation of the Receptor Tyrosine Kinase Adaptor FRS2α. In: Biochemistry. 2019 ; Vol. 58, No. 25. pp. 2809-2813.
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T1 - Myristoylation-Dependent Palmitoylation of the Receptor Tyrosine Kinase Adaptor FRS2α

AU - Barylko, Barbara

AU - Chen, Yu Ju

AU - Hennen, Jared

AU - Angert, Isaac

AU - Chen, Yan

AU - Mueller, Joachim D.

AU - Sun, Hui Qiao

AU - Taylor, Clinton A.

AU - Liou, Jen

AU - Yin, Helen

AU - Albanesi, Joseph P.

PY - 2019/6/25

Y1 - 2019/6/25

N2 - An early step in signaling from activated receptor tyrosine kinases (RTKs) is the recruitment of cytosolic adaptor proteins to autophosphorylated tyrosines in the receptor cytoplasmic domains. Fibroblast growth factor receptor substrate 2α (FRS2α) associates via its phosphotyrosine-binding domain (PTB) to FGF receptors (FGFRs). Upon FGFR activation, FRS2α undergoes phosphorylation on multiple tyrosines, triggering recruitment of the adaptor Grb2 and the tyrosine phosphatase Shp2, resulting in stimulation of PI3K/AKT and MAPK signaling pathways. FRS2α also undergoes N-myristoylation, which was shown to be important for its localization to membranes and its ability to stimulate downstream signaling events (Kouhara et al., 1997). Here we show that FRS2α is also palmitoylated in cells and that cysteines 4 and 5 account for the entire modification. We further show that mutation of those two cysteines interferes with FRS2α localization to the plasma membrane (PM), and we quantify this observation using fluorescence fluctuation spectroscopy approaches. Importantly, prevention of myristoylation by introduction of a G2A mutation also abrogates palmitoylation, raising the possibility that signaling defects previously ascribed to the G2A mutant may actually be due to a failure of that mutant to undergo palmitoylation. Our results demonstrate that FRS2α undergoes coupled myristoylation and palmitoylation. Unlike stable cotranslational modifications, such as myristoylation and prenylation, palmitoylation is reversible due to the relative lability of the thioester linkage. Therefore, palmitoylation may provide a mechanism, in addition to phosphorylation, for dynamic regulation of FRS2 and its downstream signaling pathways.

AB - An early step in signaling from activated receptor tyrosine kinases (RTKs) is the recruitment of cytosolic adaptor proteins to autophosphorylated tyrosines in the receptor cytoplasmic domains. Fibroblast growth factor receptor substrate 2α (FRS2α) associates via its phosphotyrosine-binding domain (PTB) to FGF receptors (FGFRs). Upon FGFR activation, FRS2α undergoes phosphorylation on multiple tyrosines, triggering recruitment of the adaptor Grb2 and the tyrosine phosphatase Shp2, resulting in stimulation of PI3K/AKT and MAPK signaling pathways. FRS2α also undergoes N-myristoylation, which was shown to be important for its localization to membranes and its ability to stimulate downstream signaling events (Kouhara et al., 1997). Here we show that FRS2α is also palmitoylated in cells and that cysteines 4 and 5 account for the entire modification. We further show that mutation of those two cysteines interferes with FRS2α localization to the plasma membrane (PM), and we quantify this observation using fluorescence fluctuation spectroscopy approaches. Importantly, prevention of myristoylation by introduction of a G2A mutation also abrogates palmitoylation, raising the possibility that signaling defects previously ascribed to the G2A mutant may actually be due to a failure of that mutant to undergo palmitoylation. Our results demonstrate that FRS2α undergoes coupled myristoylation and palmitoylation. Unlike stable cotranslational modifications, such as myristoylation and prenylation, palmitoylation is reversible due to the relative lability of the thioester linkage. Therefore, palmitoylation may provide a mechanism, in addition to phosphorylation, for dynamic regulation of FRS2 and its downstream signaling pathways.

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