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J Biol Chem, Vol. 274, Issue 34, 24401-24407, August 20, 1999

An Enhancer Element for Expression of the Ncx (Enx, Hox11L1) Gene in Neural Crest-derived Cells

Yoshinori Iitsuka^§, Hironori Shimizu, Myeng M. Kang, Kazushi Sasagawa, Souei Sekiya^§, Takeshi Tokuhisa, and Masahiko Hatano

From the  Department of Developmental Genetics, Chiba University Graduate School of Medicine, Chiba 260-8670, Japan and the ^§ Department of Obstetrics and Gynecology, Chiba University School of Medicine, Chiba 260-8670, Japan

The murine Ncx (Enx, Hox11L1) gene is specifically expressed in a neuronal subset of neural crest-derived tissues. In attempts to elucidate the regulatory DNA element of the tissue-specific expression, we sequenced the 5'-flanking region of the Ncx gene. The transcriptional initiation site was determined at 297 nucleotides (297) upstream from the ATG start codon (+1). A retinoic acid response element was located on the region between 1163 and 1150. Transient transfection assays with the 5'-flanking sequences fused to the luciferase gene showed that the region between 1387 and 1368 was crucial for the tissue-specific enhancer activity. Furthermore, nuclear proteins extracted from neural crest-derived cells such as murine and human neuroblastoma cells bind to the DNA region between 1387 and 1368. This DNA element was also conserved in the 5'-flanking region of the human NCX gene. Our observations strongly suggest that the DNA element (between 1387 and 1368) contributes to tissue-specific expression of the Ncx gene in murine and human species.

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