Int J Biol Sci 2013; 9(9):872-886. doi:10.7150/ijbs.7126 This issue
1. Institute of Fisheries Sciences, National Taiwan University, Taipei 106, Taiwan;
2. Institute of Biological Chemistry, Academia Sinica, Taipei 115, Taiwan;
3. Department of Biochemistry, Taipei Medical University, Taipei 110, Taiwan;
4. Institute of Cellular and Organismic Biology, Academia Sinica, Taipei 115, Taiwan.
* These authors contributed equally to this work.
The notochord is required for body plan patterning in vertebrates, and defects in notochord development during embryogenesis can lead to diseases affecting the adult. It is therefore important to elucidate the gene regulatory mechanism underlying notochord formation. In this study, we cloned the zebrafish zinc finger 219-like (ZNF219L) based on mammalian ZNF219, which contains nine C2H2-type zinc finger domains. Through whole-mount in situ hybridization, we found that znf219L mRNA is mainly expressed in the zebrafish midbrain-hindbrain boundary, hindbrain, and notochord during development. The znf219L morpholino knockdown caused partial abnormal notochord phenotype and reduced expression of endogenous col2a1a in the notochord specifically. In addition, ZNF219L could recognize binding sites with GGGGG motifs and trigger augmented activity of the col2a1a promoter in a luciferase assay. Furthermore, in vitro binding experiments revealed that ZNF219L recognizes the GGGGG motifs in the promoter region of the zebrafish col2a1a gene through its sixth and ninth zinc finger domains. Taken together, our results reveal that ZNF219L is involved in regulating the expression of col2a1a in zebrafish notochord specifically.
Keywords: zinc finger protein 219, notochord, zebrafish, collagen type 2 alpha 1a, transcriptional regulation.