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First published online May 28, 2004
doi: 10.1242/10.1242/dev.01167

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her3, a zebrafish member of the hairy-E(spl) family, is repressed by Notch signalling

Stefan Hans^1,*,¶, Nico Scheer^1,, Iris Riedl, Elisabeth v. Weizsäcker¹, Patrick Blader^2, and José A. Campos-Ortega^1,

¹ Institut für Entwicklungsbiologie, Universität zu Köln, 50923 Köln, Germany
² IGBMC, Parc d'Innovation, BP163, 67404 Illkirch Cedex, France

^¶ Authors for correspondence (e-mail: stefan{at}uoneuro.uoregon.edu and Eberhard.Rudloff{at}uni-koeln.de)

Accepted 12 March 2004

her3 encodes a zebrafish bHLH protein of the Hairy-E(Spl) family. During embryogenesis, the gene is transcribed exclusively in the developing central nervous system, according to a fairly simple pattern that includes territories in the mesencephalon/rhombencephalon and the spinal cord. In all territories, the her3 transcription domain encompasses regions in which neurogenin 1 (neurog1) is not transcribed, suggesting regulatory interactions between the two genes. Indeed, injection of her3 mRNA leads to repression of neurog1 and to a reduction in the number of primary neurones, whereas her3 morpholino oligonucleotides cause ectopic expression of neurog1 in the rhombencephalon. Fusions of Her3 to the transactivation domain of VP16 and to the repression domain of Engrailed show that Her3 is indeed a transcriptional repressor. Dissection of the Her3 protein reveals two possible mechanisms for transcriptional repression: one mediated by the bHLH domain and the C-terminal WRPW tetrapeptide; and the other involving the N-terminal domain and the orange domain. Gel retardation assays suggest that the repression of neurog1 transcription occurs by binding of Her3 to specific DNA sequences in the neurog1 promoter. We have examined interrelationships of her3 with members of the Notch signalling pathway by the Gal4-UAS technique and mRNA injections. The results indicate that Her3 represses neurog1 and, probably as a consequence of the neurog1 repression, deltaA, deltaD and her4. Moreover, Her3 represses its own transcription as well. Surprisingly, and in sharp contrast to other members of the E(spl) gene family, transcription of her3 is repressed rather than activated by Notch signalling.

Key words: Zebrafish, neurog1 (ngn1), Her3

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