Structure and Promoter Analysis of Math3 Gene, a Mouse Homolog of Drosophila Proneural Geneatonal
NEURAL-SPECIFIC EXPRESSION BY DUAL PROMOTER ELEMENTS*
- From the Department of Biological Sciences, Kyoto University Faculty of Medicine, Yoshida, Sakyo-ku, Kyoto 606, Japan
Abstract
ath3, a vertebrate basic helix-loop-helix gene homologous to Drosophila proneural gene atonal, can directly convert non-neural cells into neurons with the anterior features. In the mouse, ath3expression initially occurs widely in the developing nervous system and then gradually becomes restricted to the neural retina. Here, we characterized the genomic organization and promoter activity of mouseath3 (Math3). Math3 gene consists of two exons separated by an 8-kilobase intron, and the whole protein-coding region is located in the second exon. Transcription starts at two sites, which are 75 nucleotides apart from each other, and there is no typical TATA box in the upstream region of either start site. Transient transfection analysis showed that the 5′-region ofMath3 can direct efficient expression in neuroblastoma cells but not in glioma or fibroblast cells. Deletion studies revealed that the proximal 193-base pair region, which contains the downstream transcription initiation site but not the upstream site, is essential for the Math3 promoter activity and can direct efficient expression in neuroblastoma cells. In contrast, retrovirus-mediated promoter analysis demonstrated that a region further upstream is additionally necessary for retinal expression. These results indicate that Math3 promoter contains two essential regulatory regions, the proximal 193-base pair region, which confers efficient neural-specific expression, and a region further upstream, required for retinal expression.
Footnotes
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↵* This work was supported by research grants from the Ministry of Education, Science, and Culture of Japan and Special Coordination Funds for Promoting Science and Technology.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBank™/EMBL Data Bank with accession number(s) and .
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↵‡ Present address: The Burnham Institute, La Jolla Cancer Research Foundation, 10901 N. Torrey Pines Rd., La Jolla, CA 92037.
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↵§ To whom correspondence should be addressed: Tel.: 81-75-753-4438; Fax: 81-75-753-4404.
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↵1 The abbreviations used are: bHLH, basic helix-loop-helix; bp, base pair(s); kb, kilobase(s); INL, inner nuclear layer; Math3, mouse atonal homolog 3; ONL, outer nuclear layer; PCR, polymerase chain reaction; X-gal, 5-bromo-4-chloro-3-indolyl β-d-galactopyranoside.
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↵2 H. Tsuda, K. Takebayashi, S. Nakanishi, and R. Kageyama, unpublished data.
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- Received July 30, 1997.
- Revision received November 17, 1997.
- The American Society for Biochemistry and Molecular Biology, Inc.
