Cloning And Functional Analysis Of Novel Genes, ZNF411 And ZNF415, Expressed In Human Heart

The heart is one of first structures to form during organogenesis in the vertebrate. Cardiogenesis is a very complex process and it requires precise regulation of series of genes. The identification of the molecular pathways that guide heart morphogenesis will provide a basic understanding of how heart is formed and of how congenital heart diseases(CHD) are taken place. Zinc finger family is one of the largest humangene family, zinc finger proteins are thought to function astranscriptional regulators for nucleic acid. binding.Mitogen-activated protein kinase (MAPK) signal transduction pathways are among the most widespread mechanisms of eukaryotic cell regulation, and Many transcription factors are probably important MAPK targets. With the aim of identifying novel genes involved in human heart development and diseases, two novel genes, ZNF411 and ZNF415, were isolated from heart cDNA library. These two genes map to chromosome 19q and belong to Cys2/His2 zinc finger protein family. Northern Blot indicated that ZNF411 gene is expressed in heart and skeletal muscle, while ZNF415 is strongly expressed only in heart tissue. It suggested that these two genes may have important function in heart development and regulation. In addition, PEGFP-ZNF411 and PEGFP-ZNF415 fusion mammalian expression protein were constructed and introduced into the COS-7 cells. Observation under the fluorescent microscopy showed that both PEGFP-ZNF411 and PEGFP-ZNF415 were expressed and existed mainly in the nucleus, suggesting that ZNF411 and ZNF415 gene-coding products may be nuclear proteins. We synthesized multipeptides including 20 amino acid residues of the ZNF415 gene using Fmoc solid phase chemistry and obtained its multiclonal antibody by using rabbit immune serum, which provided facility in studyingits function by immunotochemistry. Reporter gene assays show that ZNP411 is a transcriptional repressor and overexpression of ZNF411 in the COS-7 cells inhibits the transcriptional activities of AP-1 and SRE, suggesting that ZNF411 is probably negative regulator in MARK signaling pathway. Taken together, ZNF411 and ZNF415 are regulational factors during heart development and probably are candidate genes of heart diseases.