Reciprocal regulation of Neu tyrosine kinase activity and caveolin-1 protein expression in vitro and in vivo: Implications for human breast cancer

Jeffrey A. Engelman, Richard J. Lee, Anthony Karnezis, David J. Bearss, Marc Webster, Peter Siegel, William J. Muller, Jolene J. Windle, Richard G. Pestell, Michael P. Lisanti

Research output: Contribution to journalArticle

187 Citations (Scopus)

Abstract

Neu (c-erbB2) is a proto-oncogene product that encodes an epidermal growth factor-like receptor tyrosine kinase. Amplification of wild-type c- Neu and mutational activation of Neu (Neu T) have been implicated in oncogenic transformation of cultured fibroblasts and mammary tumorigenesis in vivo. Here, we examine the relationship between Neu tyrosine kinase activity and caveolin-1 protein expression in vitro and in vivo. Recent studies have suggested that caveolins may function as negative regulators of signal transduction. Our current results show that mutational activation of c-Neu down-regulates caveolin-1 protein expression, but not caveolin-2, in cultured NIH 3T3 and Rat 1 cells. Conversely, recombinant overexpression of caveolin- 1 blocks Neu-mediated signal transduction in vivo. These results suggest a reciprocal relationship between c-Neu tyrosine kinase activity and caveolin- 1 protein expression. We next analyzed a variety of caveolin-1 deletion mutants to map this caveolin-1-dependent inhibitory activity to a given region of the caveolin-1 molecule. Results from this mutational analysis show that this functional in vivo inhibitory activity is contained within caveolin-1 residues 32-95. In accordance with these in vivo studies, a 20- amino acid peptide derived from this region (the caveolin-1 scaffolding domain) was sufficient to inhibit Neu autophosphorylation in an in vitro kinase assay. To further confirm or refute the relevance of our findings in vivo, we next examined the expression levels of caveolin-1 in mammary tumors derived from c-Neu transgenic mice. Our results indicate that dramatic reduction of caveolin-1 expression occurs in mammary tumors derived from c- Neu-expressing transgenic mice and other transgenic mice expressing downstream effectors of Neu-mediated signal transduction, such as Src and Ras. Taken together, our data suggest that a novel form of reciprocal negative regulation exists between c-Neu and caveolin-1.

Original languageEnglish (US)
Pages (from-to)20448-20455
Number of pages8
JournalJournal of Biological Chemistry
Volume273
Issue number32
DOIs
StatePublished - Aug 7 1998
Externally publishedYes

Fingerprint

Caveolins
Caveolin 1
Protein-Tyrosine Kinases
Breast Neoplasms
Proteins
Signal transduction
Transgenic Mice
Signal Transduction
In Vitro Techniques
Tumors
Caveolin 2
Chemical activation
Proto-Oncogenes
Oncogene Proteins
Receptor Protein-Tyrosine Kinases
Fibroblasts
Epidermal Growth Factor Receptor
Epidermal Growth Factor
Amplification
Rats

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Reciprocal regulation of Neu tyrosine kinase activity and caveolin-1 protein expression in vitro and in vivo : Implications for human breast cancer. / Engelman, Jeffrey A.; Lee, Richard J.; Karnezis, Anthony; Bearss, David J.; Webster, Marc; Siegel, Peter; Muller, William J.; Windle, Jolene J.; Pestell, Richard G.; Lisanti, Michael P.

In: Journal of Biological Chemistry, Vol. 273, No. 32, 07.08.1998, p. 20448-20455.

Research output: Contribution to journalArticle

Engelman, JA, Lee, RJ, Karnezis, A, Bearss, DJ, Webster, M, Siegel, P, Muller, WJ, Windle, JJ, Pestell, RG & Lisanti, MP 1998, 'Reciprocal regulation of Neu tyrosine kinase activity and caveolin-1 protein expression in vitro and in vivo: Implications for human breast cancer', Journal of Biological Chemistry, vol. 273, no. 32, pp. 20448-20455. https://doi.org/10.1074/jbc.273.32.20448
Engelman, Jeffrey A. ; Lee, Richard J. ; Karnezis, Anthony ; Bearss, David J. ; Webster, Marc ; Siegel, Peter ; Muller, William J. ; Windle, Jolene J. ; Pestell, Richard G. ; Lisanti, Michael P. / Reciprocal regulation of Neu tyrosine kinase activity and caveolin-1 protein expression in vitro and in vivo : Implications for human breast cancer. In: Journal of Biological Chemistry. 1998 ; Vol. 273, No. 32. pp. 20448-20455.
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abstract = "Neu (c-erbB2) is a proto-oncogene product that encodes an epidermal growth factor-like receptor tyrosine kinase. Amplification of wild-type c- Neu and mutational activation of Neu (Neu T) have been implicated in oncogenic transformation of cultured fibroblasts and mammary tumorigenesis in vivo. Here, we examine the relationship between Neu tyrosine kinase activity and caveolin-1 protein expression in vitro and in vivo. Recent studies have suggested that caveolins may function as negative regulators of signal transduction. Our current results show that mutational activation of c-Neu down-regulates caveolin-1 protein expression, but not caveolin-2, in cultured NIH 3T3 and Rat 1 cells. Conversely, recombinant overexpression of caveolin- 1 blocks Neu-mediated signal transduction in vivo. These results suggest a reciprocal relationship between c-Neu tyrosine kinase activity and caveolin- 1 protein expression. We next analyzed a variety of caveolin-1 deletion mutants to map this caveolin-1-dependent inhibitory activity to a given region of the caveolin-1 molecule. Results from this mutational analysis show that this functional in vivo inhibitory activity is contained within caveolin-1 residues 32-95. In accordance with these in vivo studies, a 20- amino acid peptide derived from this region (the caveolin-1 scaffolding domain) was sufficient to inhibit Neu autophosphorylation in an in vitro kinase assay. To further confirm or refute the relevance of our findings in vivo, we next examined the expression levels of caveolin-1 in mammary tumors derived from c-Neu transgenic mice. Our results indicate that dramatic reduction of caveolin-1 expression occurs in mammary tumors derived from c- Neu-expressing transgenic mice and other transgenic mice expressing downstream effectors of Neu-mediated signal transduction, such as Src and Ras. Taken together, our data suggest that a novel form of reciprocal negative regulation exists between c-Neu and caveolin-1.",
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AU - Bearss, David J.

AU - Webster, Marc

AU - Siegel, Peter

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