Department of Human Genetics, Hannover Medical School, Hannover, Germany.
Department of Pediatric Gastroenterology and Hepatology, Division of Kidney, Liver and Metabolic Diseases, Hannover Medical School, Hannover, Germany.
Ann Hum Genet. 2020 Mar;84(2):195-200. doi: 10.1111/ahg.12355. Epub 2019 Oct 9.
Wilson's disease is an autosomal recessive disorder resulting from copper excess. Some patients with clinical Wilson's disease symptoms exhibit no or only heterozygous pathogenic variants in the coding region of the disease-causing ATP7B gene. Therefore, the ATP7B promoter region is of special interest. Metal-responsive elements (MREs) located in the ATP7B promoter are promising motifs in modulating the ATP7B expression. We studied protein interaction of MREe, MREc, and MREd by electrophoretic mobility shift assays and revealed specific interactions for all MREs. We further narrowed down the specific binding site. Proteins potentially binding to the three MREs were identified by MatInspector analyses. Metal regulatory transcription factor 1 (MTF1) could be validated to bind to MREe by electrophoretic mobility shift assays. ATP7B promoter-driven reporter gene expression was significantly increased because of this interaction. MTF1 is a strong candidate in regulating the ATP7B expression through MREe binding.
威尔逊病是一种常染色体隐性遗传病，由铜过量引起。一些有临床威尔逊病症状的患者在致病 ATP7B 基因的编码区没有表现出或只有杂合的致病性变异。因此，ATP7B 启动子区域特别引人注目。位于 ATP7B 启动子中的金属反应元件（MRE）是调节 ATP7B 表达的有前途的基序。我们通过电泳迁移率变动分析研究了 MREe、MREc 和 MREd 的蛋白相互作用，并揭示了所有 MRE 的特异性相互作用。我们进一步缩小了特定的结合位点。通过 MatInspector 分析鉴定了可能与这三个 MRE 结合的蛋白质。电泳迁移率变动分析验证了金属调节转录因子 1（MTF1）可以与 MREe 结合。由于这种相互作用，ATP7B 启动子驱动的报告基因表达显著增加。MTF1 是通过 MREe 结合调节 ATP7B 表达的一个强有力的候选者。
