Identification and characterization of alternative promoters of zebrafish Rtn-4/Nogo genes in cultured cells and zebrafish embryos

Yi Chung Chen, Bo Kai Wu, Cheng Ying Chu, Chia Hsiung Cheng, Hau Wei Han, Gen Der Chen, Ming Ting Lee, Pung Pung Hwang, Koichi Kawakami, Chun Che Chang, Chang Jen Huang

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Abstract

In mammals, the Nogo family consists of Nogo-A, Nogo-B and Nogo-C. However, there are three Rtn-4/Nogo-related transcripts were identified in zebrafish. In addition to the common C-terminal region, the N-terminal regions of Rtn4-n/Nogo-C1, Rtn4-m/Nogo-C2 and Rtn4-l/Nogo-B, respectively, contain 9, 25 and 132 amino acid residues. In this study, we isolated the 50-upstream region of each gene from a BAC clone and demonstrated that the putative promoter regions, P1-P3, are functional in cultured cells and zebrafish embryos. A transgenic zebrafish Tg(Nogo-B:GFP) line was generated using P1 promoter region to drive green fluorescent protein (GFP) expression through Tol2-mediated transgenesis. This line recapitulates the endogenous expression pattern of Rtn4-l/Nogo-B mRNA in the brain, brachial arches, eyes, muscle, liver and intestines. In contrast, GFP expressions by P2 and P3 promoters were localized to skeletal muscles of zebrafish embryos. Several GATA and E-box motifs are found in these promoter regions. Using morpholino knockdown experiments, GATA4 and GATA6 were involved in the control of P1 promoter activity in the liver and intestine, while Myf5 and MyoD for the control of P1 and P3 promoter activities in muscles. These data demonstrate that zebrafish Rtn4/Nogo transcripts might be generated by coupling mechanisms of alternative first exons and alternative promoter usage.

Original languageEnglish
Article numbergkq230
Pages (from-to)4635-4650
Number of pages16
JournalNucleic Acids Research
Volume38
Issue number14
DOIs
Publication statusPublished - Apr 8 2010

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ASJC Scopus subject areas

  • Genetics
  • Medicine(all)

Cite this

Chen, Y. C., Wu, B. K., Chu, C. Y., Cheng, C. H., Han, H. W., Chen, G. D., Lee, M. T., Hwang, P. P., Kawakami, K., Chang, C. C., & Huang, C. J. (2010). Identification and characterization of alternative promoters of zebrafish Rtn-4/Nogo genes in cultured cells and zebrafish embryos. Nucleic Acids Research, 38(14), 4635-4650. [gkq230]. https://doi.org/10.1093/nar/gkq230