Department of Physiology and Neurobiology, University of Connecticut, Storrs, Connecticut, United States of America.
PLoS One. 2012;7(2):e32404. doi: 10.1371/journal.pone.0032404. Epub 2012 Feb 27.
ZNF804a was identified by a genome-wide association study (GWAS) in which a single nucleotide polymorphism (SNP rs1344706) in ZNF804a reached genome-wide statistical significance for association with a combined diagnosis of schizophrenia (SZ) and bipolar disorder. Although the molecular function of ZNF804a is unknown, the amino acid sequence is predicted to contain a C2H2-type zinc-finger domain and suggests ZNF804a plays a role in DNA binding and transcription. Here, we confirm that ZNF804a directly contributes to transcriptional control by regulating the expression of several SZ associated genes and directly interacts with chromatin proximal to the promoter regions of PRSS16 and COMT, the two genes we find upregulated by ZNF804a. Using immunochemistry we establish that ZNF804a is localized to the nucleus of rat neural progenitor cells in culture and in vivo. We demonstrate that expression of ZNF804a results in a significant increase in transcript levels of PRSS16 and COMT, relative to GFP transfected controls, and a statistically significant decrease in transcript levels of PDE4B and DRD2. Furthermore, we show using chromatin immunoprecipitation assays (ChIP) that both epitope-tagged and endogenous ZNF804a directly interacts with the promoter regions of PRSS16 and COMT, suggesting a direct upregulation of transcription by ZNF804a on the expression of these genes. These results are the first to confirm that ZNF804a regulates transcription levels of four SZ associated genes, and binds to chromatin proximal to promoters of two SZ genes. These results suggest a model where ZNF804a may modulate a transcriptional network of SZ associated genes.
锌指蛋白 804a（ZNF804a）是通过全基因组关联研究（GWAS）鉴定的，该研究中 ZNF804a 中的一个单核苷酸多态性（SNP rs1344706）与精神分裂症（SZ）和双相情感障碍的综合诊断达到了全基因组统计学意义上的关联。虽然 ZNF804a 的分子功能未知，但氨基酸序列预计包含一个 C2H2 型锌指结构域，表明 ZNF804a 在 DNA 结合和转录中发挥作用。在这里，我们通过调节几个与 SZ 相关基因的表达，证实 ZNF804a 直接参与转录控制，并直接与 PRSS16 和 COMT 启动子区域附近的染色质相互作用，我们发现这两个基因被 ZNF804a 上调。我们使用免疫化学方法确定 ZNF804a 在培养的大鼠神经祖细胞和体内定位于细胞核。我们证明，与 GFP 转染对照相比，ZNF804a 的表达导致 PRSS16 和 COMT 的转录本水平显著增加，而 PDE4B 和 DRD2 的转录本水平显著降低。此外，我们通过染色质免疫沉淀测定（ChIP）显示，表位标记和内源性 ZNF804a 都直接与 PRSS16 和 COMT 的启动子区域相互作用，表明 ZNF804a 直接上调这些基因的转录。这些结果是第一个证实 ZNF804a 调节四个与 SZ 相关基因的转录水平，并与两个 SZ 基因的启动子附近染色质结合的研究结果。这些结果表明，ZNF804a 可能调节与 SZ 相关基因的转录网络的模型。
