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Molecular and Cellular Biology, June 2004, p. 5257-5268, Vol. 24, No. 12
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.12.5257-5268.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Characterization of NF-B/IB Proteins in Zebra Fish and Their Involvement in Notochord Development

The Salk Institute for Biological Studies, La Jolla, California 92037-1099

Received 29 January 2004/ Accepted 12 March 2004

Although largely involved in innate and adaptive immunity, NF-B plays an important role in vertebrate development. In chicks, the inactivation of the NF-B pathway induces functional alterations of the apical ectodermal ridge, which mediates limb outgrowth. In mice, the complete absence of NF-B activity leads to prenatal death and neural tube defects. Here, we report the cloning and characterization of NF-B/IB proteins in zebra fish. Despite being ubiquitously expressed among the embryonic tissues, NF-B/IB members present distinct patterns of gene expression during the early zebra fish development. Biochemical assays indicate that zebra fish NF-B proteins are able to bind consensus DNA-binding (B) sites and inhibitory IB proteins from mammals. We show that zebra fish IBs are degraded in a time-dependent manner after induction of transduced murine embryo fibroblasts (MEFs) and that these proteins are able to rescue NF-B activity in IB^–/– MEFs. Expression of a dominant-negative form of the murine IB (mIBM), which is able to block NF-B in zebra fish cells, interferes with the notochord differentiation, generating no tail (ntl)-like embryos. This phenotype can be rescued by coinjection of the T-box gene ntl (Brachyury homologue), which is typically required for the formation of posterior mesoderm and axial development, suggesting that ntl lies downstream of NF-B. We further show that ntl and Brachyury promoter regions contain functional B sites and NF-B can directly modulate ntl expression. Our study illustrates the conservation and compatibility of NF-B/IB proteins among vertebrates and the importance of NF-B pathway in mesoderm formation during early embryogenesis.

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