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Vol. 15, No. 22, pp. 3039-3049, November 15, 2001

RESEARCH PAPER
Role of Dlx6 in regulation of an endothelin-1-dependent, dHAND branchial arch enhancer

Jeroen Charité,1,7 David G. McFadden,1 Giorgio Merlo,5 Giovanni Levi,5,6 David E. Clouthier,2,8 Masashi Yanagisawa,2,4 James A. Richardson,3 and Eric N. Olson1,9

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]


Present addresses: 7Department of Cell Biology and Genetics, Erasmus University Rotterdam, Dr. Molewaterplein 50, 3015GE Rotterdam, The Netherlands; 8Department of Molecular, Cellular and Craniofacial Biology, Birth Defects Center, School of Dentistry, University of Louisville, 501 S. Preston St., Louisville, KY 40292, USA.

9 Corresponding author.


GENES & DEVELOPMENT 15:3039-3049 © 2001 by Cold Spring Harbor Laboratory Press  ISSN 0890-9369/01 $5.00

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