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J Biol Chem, Vol. 273, Issue 32, 20448-20455, August 7, 1998
From the a Department of Molecular Pharmacology and the
c Departments of Developmental and Molecular Biology and
Medicine, Albert Einstein Cancer Center, Albert Einstein College of
Medicine, Bronx, New York 10461, the e Cancer Therapy and
Research Center, San Antonio, Texas 78229, and the
g Department of Pathology, McMaster University, West
Hamilton, Ontario L8S 4K1, Canada
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.
Reciprocal Regulation of Neu Tyrosine Kinase Activity and
Caveolin-1 Protein Expression in Vitro and in
Vivo
IMPLICATIONS FOR HUMAN BREAST CANCER
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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