Abstract
BackgroundZNF143 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.ResultsThe 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.ConclusionsNormal 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.
Highlights
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
To verify the mRNA gene activating potential of zebrafish ZNF143 and demarcate boundaries of this region, we fused fragments encoding zebrafish ZNF143 to the S. cerevisiae GAL4p DNA binding domain, and performed transient transfection assays with such expression plasmids and a firefly luciferase reporter gene transcribed from a minimal promoter driven by GAL4 binding sites
The zebrafish pax2a gene promoter contains binding sites for ZNF143 and is controlled by this protein in transfected cells Since pax2a gene expression was affected after ZNF143 knockdown, we investigated whether this transcription factor might control zebrafish pax2a transcription
Summary
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. The vertebrate transcriptional activator protein, ZNF143 ( known as STAF for selenocysteine tRNA gene transcription activating factor, or SBF for SPH-binding factor) operates at a multitude of small RNA and protein-coding gene promoters [1,2,3,4,5]. Because of the highly degenerate and relatively long DNA-binding site recognized by ZNF143, it was not recognized for many years that approximately 2000 mammalian protein-coding genes contain SPH (SphI Postoctamer Homology [16]) elements, or STAF Binding Sites (SBS), in their promoters [5]. Many potential small RNA and protein-coding gene promoters are targeted, but which are most pivotal in vivo?
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