1. Institute of Fisheries Sciences, National Taiwan University, Taipei 106, Taiwan;
Int J Biol Sci. 2013 Sep 5;9(9):872-86. doi: 10.7150/ijbs.7126. eCollection 2013.
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.
脊索是脊椎动物体节模式形成所必需的,胚胎发生过程中脊索发育缺陷可导致影响成年的疾病。因此,阐明脊索形成的基因调控机制非常重要。在这项研究中,我们基于哺乳动物 ZNF219 克隆了斑马鱼锌指 219 样(ZNF219L),它包含九个 C2H2 型锌指结构域。通过整体原位杂交,我们发现 znf219L mRNA 在发育过程中主要在斑马鱼中脑-后脑边界、后脑和脊索中表达。znf219L 型 morpholino 敲低导致部分异常脊索表型,并特异性降低脊索中的内源性 col2a1a 表达。此外,ZNF219L 可以识别具有 GGGGG 基序的结合位点,并在荧光素酶测定中触发 col2a1a 启动子的增强活性。此外,体外结合实验表明,ZNF219L 通过其第六和第九个锌指结构域识别斑马鱼 col2a1a 基因启动子区域中的 GGGGG 基序。总之,我们的结果表明,ZNF219L 特异性参与调节斑马鱼脊索中 col2a1a 的表达。