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Vol. 15, No. 22, pp. 3039-3049, November 15, 2001
1 Department of Molecular Biology, 2 Department of
Molecular Genetics, 3 Department of Pathology, and
4 Howard Hughes Medical Institute, University of Texas
Southwestern Medical Center, Dallas, Texas 75239-9148, USA;
5 Laboratory of Molecular Morphogenesis, National Cancer
Institute-I.S.T., 16132 Genova, Italy; 6 Laboratoire de
Physiologie, Museum National d'Histoire Naturelle, UMR CNRS 8572, Paris, France
Neural crest cells play a key role in craniofacial development. The
endothelin family of secreted polypeptides regulates development of
several neural crest sublineages, including the branchial arch neural
crest. The basic helix-loop-helix transcription factor dHAND is also
required for craniofacial development, and in endothelin-1 (ET-1) mutant embryos, dHAND expression in the
branchial arches is down-regulated, implicating it as a transcriptional
effector of ET-1 action. To determine the mechanism that links ET-1
signaling to dHAND transcription, we analyzed the dHAND
gene for cis-regulatory elements that control transcription in
the branchial arches. We describe an evolutionarily conserved
dHAND enhancer that requires ET-1 signaling for activity. This
enhancer contains four homeodomain binding sites that are required for
branchial arch expression. By comparing protein binding to these sites
in branchial arch extracts from endothelin receptor A
(EdnrA) mutant and wild-type mouse embryos, we identified Dlx6,
a member of the Distal-less family of homeodomain proteins, as an
ET-1-dependent binding factor. Consistent with this conclusion,
Dlx6 was down-regulated in branchial arches from EdnrA
mutant mice. These results suggest that Dlx6 acts as an intermediary
between ET-1 signaling and dHAND transcription during
craniofacial morphogenesis.
[Key Words: dHAND/HAND2; neural crest; branchial arches; endothelin-1; endothelin receptor A; Dlx6]
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