Mutations in the EYA1 gene are responsible for branchio-oto-renal (BOR) syndrome as well as for other ocular defects. Most of the mutations are located within or in the vicinity of the EYA domain, which is highly conserved in the EYA protein family. The EYA domain is required for protein-protein interactions, which are important to the biological function of EYA proteins. To determine how EYA1 mutations cause BOR syndrome and/or ocular defects, we tested the effects of Eya1 mutations on interactions with Six. Dach, and G proteins by mammalian two-hybrid and GST-pulldown assays. Defective interactions were noted between BOR-type mutations S486P and L504R of Eya1 and Dach1, G proteins, and some Six proteins. These mutations impaired the activation of transcription from a Six-responsive gene, myogenin, with Six5. S486P and L504R showed an altered digestion pattern with trypsin, and L504R also decreased the sensitivity to V8 protease digestion and produced a peptide fragment with a different M(r). Our results suggest that defective protein-protein interactions of the mutations in the EYA domain underlie BOR syndrome and that SIX, DACH, and/or G proteins are possibly involved in the pathogenic processes.

中文翻译：

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患有鼠耳肾综合征患者的小鼠Eyal窝藏突变与Six，Dach和G蛋白之间的相互作用受损。

EYA1基因的突变是导致分支-耳-肾（BOR）综合征以及其他眼部缺陷的原因。大多数突变位于EYA结构域内或附近，在EYA蛋白家族中高度保守。蛋白质相互作用需要EYA结构域，这对EYA蛋白质的生物学功能很重要。为了确定EYA1突变如何导致BOR综合征和/或眼部缺陷，我们测试了Eya1突变对与6相互作用的影响。通过哺乳动物的双杂交和GST-pulldown检测得出Dach和G蛋白。Eya1和Dach1，G蛋白和一些6个蛋白的BOR型突变S486P和L504R之间存在相互作用。这些突变削弱了六反应基因，肌生成素与Six5的转录激活。S486P和L504R显示出用胰蛋白酶改变的消化模式，而L504R也降低了对V8蛋白酶消化的敏感性，并产生了具有不同M（r）的肽片段。我们的结果表明，EYA域中突变的有缺陷的蛋白质-蛋白质相互作用是BOR综合征的基础，而SIX，DACH和/或G蛋白质可能与致病过程有关。