A novel zinc finger protein 219-like (ZNF219L) is involved in the regulation of zebrafish notochord related genes expression
Date Issued
2013
Date
2013
Author(s)
Lien, Huang-Wei
Abstract
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.
On the other hand, zebrafish synuclein-γ2 (sncgb) has been reported to be expressed specifically in the notochord. However, the mechanism by which the sncgb gene promoter is regulated has not been described. In this part, we demonstrate that sncgb gene was another target gene regulated by ZNF219L. ZNF219L and sox9a are involved in the regulation of sncgb gene expression. Furthermore, we observed that over-expression of both ZNF219L and Sox9a resulted in increased sncgb expression. In addition, ZNF219L is physically associated with Sox9a, and simultaneous morpholino knockdown of znf219L and sox9a caused a synergistic decrease of sncgb expression in the notochord. Thus, these results reveal that coordination of ZNF219L with Sox9a is involved in the regulation of notochord-specific expression of sncgb.
Taken together, ZNF219L is involved in the regulation of zebrafish notochord related genes expression and our studies provided better understanding of the mechanism of notochord related genes regulation in early developmental stage.
Subjects
鋅指蛋白219
脊索
斑馬魚
第二型膠元蛋白α1a
Type
thesis
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