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J Biol Chem, Vol. 274, Issue 45, 32258-32264, November 5, 1999
-mediated Signaling Pathways
From the Ludwig Institute for Cancer Research, Post Office, Royal
Melbourne Hospital, Victoria 3050, Australia
Smad7 has been identified as a negative regulator
of transforming growth factor 
(TGF-) signaling by interfering
with the phosphorylation of other Smad proteins by TGF- receptor
type I (TRI). We established a mink lung epithelial (Mv1Lu) cell
line where ectopic expression of Smad7 is tightly controlled by
doxycycline using an improved Tet-on system. Once induced by
doxycycline, the recombinant Smad7 was localized predominantly in the
perinuclear region and in the cytoplasm. However, the type of culture
surface alters the subcellular localization of Smad7: on plastic or on fibronectin-coated glass, Smad7 was localized in the cytoplasm; but
when the cells were cultured on glass, nuclear localization was
observed. TGF- stimulation did not alter substantially the cellular
distribution of Smad7. Importantly, the expression of recombinant Smad7
differentially inhibited TGF- signaling pathways. Consistent with
previous studies, Smad7 inhibited TGF--stimulated induction of type
1 plasminogen activator inhibitor as measured by p3TP-Lux reporter.
However, expression of Smad7 had little effect on TGF--induced
growth inhibition.
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