Constitutive and growth factor-regulated phosphorylation of caveolin-1 occurs at the same site (Tyr-14) in vivo: Identification of a c-Src/Cav-1/Grb7 signaling cassette

H. Lee, D. Volonte, F. Galbiati, P. Iyengar, D. M. Lublin, D. B. Bregman, M. T. Wilson, R. Campos-Gonzalez, B. Bouzahzah, R. G. Pestell, P. E. Scherer, M. P. Lisanti

Research output: Contribution to journalArticle

272 Citations (Scopus)

Abstract

Caveolin-1 was first identified as a phosphoprotein in Rous sarcoma virus (RSV)-transformed chicken embryo fibroblasts. Tyrosine 14 is now thought to be the principal site for recognition by c-Src kinase; however, little is known about this phosphorylation event. Here, we generated a monoclonal antibody (mAb) probe that recognizes only tyrosine 14-phosphorylated caveolin-1. Using this approach, we show that caveolin-1 (Y14) is a specific tyrosine kinase substrate that is constitutively phosphorylated in Src- and Abl-transformed cells and transiently phosphorylated in a regulated fashion during growth factor signaling. We also provide evidence that tyrosine-phosphorylated caveolin-1 is localized at the major sites of tyrosine-kinase signaling, i.e. focal adhesions. By analogy with other signaling events, we hypothesized that caveolin-1 could serve as a docking site for pTyr-binding molecules. In support of this hypothesis, we show that phosphorylation of caveolin-1 on tyrosine 14 confers binding to Grb7 (an SH2-domain containing protein) both in vitro and in vivo. Furthermore, we demonstrate that binding of Grb7 to tyrosine 14-phosphorylated caveolin-1 functionally augments anchorage-independent growth and epidermal growth factor (EGF)-stimulated cell migration. We discuss the possible implications of our findings in the context of signal transduction.

Original languageEnglish
Pages (from-to)1750-1775
Number of pages26
JournalMolecular Endocrinology
Volume14
Issue number11
DOIs
Publication statusPublished - 2000 Jan 1

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Caveolin 1
Intercellular Signaling Peptides and Proteins
Phosphorylation
Tyrosine
Protein-Tyrosine Kinases
Rous sarcoma virus
Focal Adhesions
src Homology Domains
Phosphoproteins
Epidermal Growth Factor
Cell Movement
Chickens
Signal Transduction
Embryonic Structures
Fibroblasts
Binding Sites
Monoclonal Antibodies
Growth

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Endocrinology

Cite this

Lee, H. ; Volonte, D. ; Galbiati, F. ; Iyengar, P. ; Lublin, D. M. ; Bregman, D. B. ; Wilson, M. T. ; Campos-Gonzalez, R. ; Bouzahzah, B. ; Pestell, R. G. ; Scherer, P. E. ; Lisanti, M. P. / Constitutive and growth factor-regulated phosphorylation of caveolin-1 occurs at the same site (Tyr-14) in vivo : Identification of a c-Src/Cav-1/Grb7 signaling cassette. In: Molecular Endocrinology. 2000 ; Vol. 14, No. 11. pp. 1750-1775.
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abstract = "Caveolin-1 was first identified as a phosphoprotein in Rous sarcoma virus (RSV)-transformed chicken embryo fibroblasts. Tyrosine 14 is now thought to be the principal site for recognition by c-Src kinase; however, little is known about this phosphorylation event. Here, we generated a monoclonal antibody (mAb) probe that recognizes only tyrosine 14-phosphorylated caveolin-1. Using this approach, we show that caveolin-1 (Y14) is a specific tyrosine kinase substrate that is constitutively phosphorylated in Src- and Abl-transformed cells and transiently phosphorylated in a regulated fashion during growth factor signaling. We also provide evidence that tyrosine-phosphorylated caveolin-1 is localized at the major sites of tyrosine-kinase signaling, i.e. focal adhesions. By analogy with other signaling events, we hypothesized that caveolin-1 could serve as a docking site for pTyr-binding molecules. In support of this hypothesis, we show that phosphorylation of caveolin-1 on tyrosine 14 confers binding to Grb7 (an SH2-domain containing protein) both in vitro and in vivo. Furthermore, we demonstrate that binding of Grb7 to tyrosine 14-phosphorylated caveolin-1 functionally augments anchorage-independent growth and epidermal growth factor (EGF)-stimulated cell migration. We discuss the possible implications of our findings in the context of signal transduction.",
author = "H. Lee and D. Volonte and F. Galbiati and P. Iyengar and Lublin, {D. M.} and Bregman, {D. B.} and Wilson, {M. T.} and R. Campos-Gonzalez and B. Bouzahzah and Pestell, {R. G.} and Scherer, {P. E.} and Lisanti, {M. P.}",
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Lee, H, Volonte, D, Galbiati, F, Iyengar, P, Lublin, DM, Bregman, DB, Wilson, MT, Campos-Gonzalez, R, Bouzahzah, B, Pestell, RG, Scherer, PE & Lisanti, MP 2000, 'Constitutive and growth factor-regulated phosphorylation of caveolin-1 occurs at the same site (Tyr-14) in vivo: Identification of a c-Src/Cav-1/Grb7 signaling cassette', Molecular Endocrinology, vol. 14, no. 11, pp. 1750-1775. https://doi.org/10.1210/mend.14.11.0553

Constitutive and growth factor-regulated phosphorylation of caveolin-1 occurs at the same site (Tyr-14) in vivo : Identification of a c-Src/Cav-1/Grb7 signaling cassette. / Lee, H.; Volonte, D.; Galbiati, F.; Iyengar, P.; Lublin, D. M.; Bregman, D. B.; Wilson, M. T.; Campos-Gonzalez, R.; Bouzahzah, B.; Pestell, R. G.; Scherer, P. E.; Lisanti, M. P.

In: Molecular Endocrinology, Vol. 14, No. 11, 01.01.2000, p. 1750-1775.

Research output: Contribution to journalArticle

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T1 - Constitutive and growth factor-regulated phosphorylation of caveolin-1 occurs at the same site (Tyr-14) in vivo

T2 - Identification of a c-Src/Cav-1/Grb7 signaling cassette

AU - Lee, H.

AU - Volonte, D.

AU - Galbiati, F.

AU - Iyengar, P.

AU - Lublin, D. M.

AU - Bregman, D. B.

AU - Wilson, M. T.

AU - Campos-Gonzalez, R.

AU - Bouzahzah, B.

AU - Pestell, R. G.

AU - Scherer, P. E.

AU - Lisanti, M. P.

PY - 2000/1/1

Y1 - 2000/1/1

N2 - Caveolin-1 was first identified as a phosphoprotein in Rous sarcoma virus (RSV)-transformed chicken embryo fibroblasts. Tyrosine 14 is now thought to be the principal site for recognition by c-Src kinase; however, little is known about this phosphorylation event. Here, we generated a monoclonal antibody (mAb) probe that recognizes only tyrosine 14-phosphorylated caveolin-1. Using this approach, we show that caveolin-1 (Y14) is a specific tyrosine kinase substrate that is constitutively phosphorylated in Src- and Abl-transformed cells and transiently phosphorylated in a regulated fashion during growth factor signaling. We also provide evidence that tyrosine-phosphorylated caveolin-1 is localized at the major sites of tyrosine-kinase signaling, i.e. focal adhesions. By analogy with other signaling events, we hypothesized that caveolin-1 could serve as a docking site for pTyr-binding molecules. In support of this hypothesis, we show that phosphorylation of caveolin-1 on tyrosine 14 confers binding to Grb7 (an SH2-domain containing protein) both in vitro and in vivo. Furthermore, we demonstrate that binding of Grb7 to tyrosine 14-phosphorylated caveolin-1 functionally augments anchorage-independent growth and epidermal growth factor (EGF)-stimulated cell migration. We discuss the possible implications of our findings in the context of signal transduction.

AB - Caveolin-1 was first identified as a phosphoprotein in Rous sarcoma virus (RSV)-transformed chicken embryo fibroblasts. Tyrosine 14 is now thought to be the principal site for recognition by c-Src kinase; however, little is known about this phosphorylation event. Here, we generated a monoclonal antibody (mAb) probe that recognizes only tyrosine 14-phosphorylated caveolin-1. Using this approach, we show that caveolin-1 (Y14) is a specific tyrosine kinase substrate that is constitutively phosphorylated in Src- and Abl-transformed cells and transiently phosphorylated in a regulated fashion during growth factor signaling. We also provide evidence that tyrosine-phosphorylated caveolin-1 is localized at the major sites of tyrosine-kinase signaling, i.e. focal adhesions. By analogy with other signaling events, we hypothesized that caveolin-1 could serve as a docking site for pTyr-binding molecules. In support of this hypothesis, we show that phosphorylation of caveolin-1 on tyrosine 14 confers binding to Grb7 (an SH2-domain containing protein) both in vitro and in vivo. Furthermore, we demonstrate that binding of Grb7 to tyrosine 14-phosphorylated caveolin-1 functionally augments anchorage-independent growth and epidermal growth factor (EGF)-stimulated cell migration. We discuss the possible implications of our findings in the context of signal transduction.

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