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J. Biol. Chem., Vol. 275, Issue 42, 32837-32845, October 20, 2000
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From the Signal Transduction Laboratory, Institute of Molecular and
Cell Biology, 30 Medical Dr.,
Singapore 117609, Republic of Singapore
Sprouty (Spry) was first identified in a genetic
screen in Drosophila to be an antagonist of fibroblast
growth factor and epidermal growth factor (EGF) signaling, seemingly by
inhibiting the Ras/MAP kinase pathway. Data base searches lead to the
identification and cloning of, to date, four mammalian
sprouty genes. The primary sequences of the mammalian
sprouty gene products share a well conserved cysteine-rich C-terminal
domain with the Drosophila protein. The N-terminal regions,
however, do not exhibit significant homology. This study aimed at
determining the disposition of Spry proteins in intact cells before and
after stimulation of the EGF receptor tyrosine kinase. Full-length or
deletion mutants of Spry, tagged at the N termini with the
FLAG-epitope, were expressed in COS-1 cells by transient transfection
and analyzed by immunofluorescence microscopy before and after EGF
stimulation of the cells. In unstimulated cells, the Spry proteins were
distributed throughout the cytosol except for human Sprouty2 (hSpry2),
which, although generally located in the cytosol, co-localized with
microtubules. In all cases, the Spry proteins underwent rapid
translocation to membrane ruffles following EGF stimulation. The
optimal translocation domain was identified by deletion and
immunofluorescence analysis to be a highly conserved 105-amino acid
domain in the C-terminal half of the hSpry2 protein. The translocation
of this conserved domain, based on hSpry2 data, was independent of the
activation of phosphatidylinositol-3 kinase.
Sprouty Proteins Are Targeted to Membrane Ruffles upon Growth
Factor Receptor Tyrosine Kinase Activation
IDENTIFICATION OF A NOVEL TRANSLOCATION DOMAIN*
*
The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed. Tel.: 65-874-3737;
Fax: 65-779-1117; E-mail: mcbgg@imcb.nus.edu.sg.
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