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J. Biol. Chem., Vol. 276, Issue 21, 18216-18222, May 25, 2001
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From the Laboratory of Genetics, University of Wisconsin, Madison,
Wisconsin 53706
Signaling by decapentaplegic (Dpp), a
Drosophila member of the transforming growth factor (TGF)
Repression of Dpp Targets by Binding of Brinker to Mad Sites*
, and
superfamily of growth factors, has recently been shown to activate
targets such as vestigial (vg) indirectly
through negative regulation of brinker (brk). Here we show that the Brk protein functions as a repressor by binding
to Dpp response elements. The Brk DNA binding activity was localized to
an amino-terminal region containing a putative homeodomain. Brk bound
to a Dpp response element of the Ultrabithorax (Ubx) midgut enhancer at a sequence that overlaps a binding
site for the Smad protein, Mothers Against Dpp (Mad). Furthermore, Brk
was able to compete with Mad for occupancy of this binding site. This
recognition of overlapping binding sites provides a potential
explanation for why the G/C-rich Mad binding site consensus differs the
Smad3/Smad4 binding site consensus. We also found that the Dpp response
element from Ubx was more sensitive than the vg
quadrant enhancer to repression by Brk. This difference correlates with
short-range activation of Ubx by Dpp in the visceral mesoderm, whereas vg exhibits a long-range response to Dpp
in the wing imaginal disc, indicating that Brk binding sites may play a
critical role in limiting thresholds for activation by Dpp.
Finally, we provide evidence that Brk is capable of functioning as an active repressor. Thus, whereas Brk and Mad compete for regulation of Ubx and vg, Brk may regulate
other Dpp targets without direct involvement of Mad.
*
This work was supported by a grant from the National Science
Foundation (to A. L.).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.
Current address: Dept. of Pharmacology, University of Wisconsin,
Medical Sciences Center, 1300 University Ave., Madison, WI 53706.
§
To whom correspondence should be addressed. Tel.: 608-262-2456;
Fax: 608-262-2976; E-mail: alaughon@facstaff.wisc.edu.
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