Department of Pediatrics, The First Affiliated Hospital, Nanjing Medical University, Nanjing, 210029, Jiangsu, China.
Mol Biol Rep. 2020 Nov;47(11):8419-8427. doi: 10.1007/s11033-020-05882-w. Epub 2020 Oct 8.
AarF domain containing kinase 4 (ADCK4) is identified as a candidate gene associated with hereditary nephrotic syndrome (NS). Kruppel-like factor 5 (KLF5) is reported to promote podocyte survival by blocking the ERK/p38 MAPK pathways. Both ADCK4 and KLF5 are involved in the occurrence and development of podocyte disease, but their interaction remains unclear. Firstly, we found that the mRNA levels of ADCK4 and KLF5 decreased in NS patients, and both levels showed an obvious linear relationship. Secondly, we cloned the ADCK4 promoter region and examined its promoter activity in Hela, A549, and HEK 293 cell lines. Deletion analysis showed that the region - 116/- 4 relative to the transcriptional start site (TSS) was the core region of ADCK4 promoter. Thirdly, mutation analysis showed that putative binding sites for KLF5 contributed to the ADCK4 promoter activity. In HEK293 cells, we found that KLF5 upregulated the mRNA and protein levels of ADCK4. Finally, our chromatin immunoprecipitation assay found that KLF5 could bind to the specific region of ADCK4 promoter. These results showed that KLF5 can positively regulate the transcriptional activity of ADCK4.
AarF 结构域包含激酶 4（ADCK4）被鉴定为与遗传性肾病综合征（NS）相关的候选基因。Kruppel 样因子 5（KLF5）据报道通过阻断 ERK/p38 MAPK 通路促进足细胞存活。ADCK4 和 KLF5 均参与足细胞疾病的发生和发展，但它们的相互作用尚不清楚。首先，我们发现 NS 患者的 ADCK4 和 KLF5 mRNA 水平降低，且两者水平呈明显线性关系。其次，我们克隆了 ADCK4 启动子区域，并在 Hela、A549 和 HEK 293 细胞系中检测了其启动子活性。缺失分析表明，转录起始位点（TSS）前 -116/-4 为 ADCK4 启动子的核心区域。第三，突变分析表明，KLF5 的假定结合位点有助于 ADCK4 启动子活性。在 HEK293 细胞中，我们发现 KLF5 上调了 ADCK4 的 mRNA 和蛋白水平。最后，我们的染色质免疫沉淀试验发现 KLF5 可以结合到 ADCK4 启动子的特定区域。这些结果表明 KLF5 可以正向调节 ADCK4 的转录活性。
