Department of Biochemistry and Biophysics, Texas A&M University, College Station, TX 77843-2128, USA.
BMC Mol Biol. 2012 Jan 23;13:3. doi: 10.1186/1471-2199-13-3.
ZNF143 is a sequence-specific DNA-binding protein that stimulates transcription of both small RNA genes by RNA polymerase II or III, or protein-coding genes by RNA polymerase II, using separable activating domains. We describe phenotypic effects following knockdown of this protein in developing Danio rerio (zebrafish) embryos by injection of morpholino antisense oligonucleotides that target znf143 mRNA.
The loss of function phenotype is pleiotropic and includes a broad array of abnormalities including defects in heart, blood, ear and midbrain hindbrain boundary. Defects are rescued by coinjection of synthetic mRNA encoding full-length ZNF143 protein, but not by protein lacking the amino-terminal activation domains. Accordingly, expression of several marker genes is affected following knockdown, including GATA-binding protein 1 (gata1), cardiac myosin light chain 2 (cmlc2) and paired box gene 2a (pax2a). The zebrafish pax2a gene proximal promoter contains two binding sites for ZNF143, and reporter gene transcription driven by this promoter in transfected cells is activated by this protein.
Normal development of zebrafish embryos requires ZNF143. Furthermore, the pax2a gene is probably one example of many protein-coding gene targets of ZNF143 during zebrafish development.
ZNF143 是一种序列特异性 DNA 结合蛋白，它使用可分离的激活结构域，刺激 RNA 聚合酶 II 或 III 转录小 RNA 基因，或 RNA 聚合酶 II 转录蛋白编码基因。我们通过注射靶向 znf143 mRNA 的 morpholino 反义寡核苷酸，在发育中的 Danio rerio（斑马鱼）胚胎中敲低这种蛋白质，描述了随后出现的表型效应。
功能丧失表型是多效的，包括广泛的异常，包括心脏、血液、耳朵和中脑后脑边界的缺陷。通过共注射编码全长 ZNF143 蛋白的合成 mRNA 可以挽救缺失功能表型，但不能通过缺乏氨基末端激活结构域的蛋白来挽救。因此，在敲低后，包括 GATA 结合蛋白 1（gata1）、肌球蛋白轻链 2（cmlc2）和配对盒基因 2a（pax2a）在内的几个标记基因的表达受到影响。斑马鱼 pax2a 基因近端启动子包含两个 ZNF143 的结合位点，并且由该启动子驱动的转染细胞中的报告基因转录被该蛋白激活。
斑马鱼胚胎的正常发育需要 ZNF143。此外，pax2a 基因可能是 ZNF143 在斑马鱼发育过程中许多蛋白编码基因靶标的一个例子。
