Mutations in EBF3 Disturb Transcriptional Profiles and Cause Intellectual Disability, Ataxia, and Facial Dysmorphism.
Mutations in EBF3 Disturb Transcriptional Profiles and Cause Intellectual Disability, Ataxia, and Facial Dysmorphism.
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DOI：
10.1016/j.ajhg.2016.11.012
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发表时间：
2017-01-05
影响因子：
9.8
通讯作者：
Kutsche K
中科院分区：
文献类型：
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作者：
Harms FL;Girisha KM;Hardigan AA;Kortüm F;Shukla A;Alawi M;Dalal A;Brady L;Tarnopolsky M;Bird LM;Ceulemans S;Bebin M;Bowling KM;Hiatt SM;Lose EJ;Primiano M;Chung WK;Juusola J;Akdemir ZC;Bainbridge M;Charng WL;Drummond-Borg M;Eldomery MK;El-Hattab AW;Saleh MAM;Bézieau S;Cogné B;Isidor B;Küry S;Lupski JR;Myers RM;Cooper GM;Kutsche K
通过一项基于genematcher的国际合作，我们确定了10名受智力残疾、语言延迟、共济失调和面部畸形影响的个体，并通过全外显子组测序检测到携带有害的EBF3变体。在EBF3中发现1个9-bp重复和1个剪接位点、5个错义和2个无义变异；突变发生在8个个体中，错义变体c.625C >t （p.a g209trp）由两个受影响的兄弟姐妹从其健康的马赛克母亲那里遗传。EBF3属于早期B细胞因子家族（也称为Olf、COE或O/E），是一种参与神经元分化和成熟的转录因子。结构分析预测，这5个氨基酸的取代对EBF3的DNA结合具有破坏作用。在HEK293T细胞中短暂表达EBF3突变蛋白，发现细胞质中除了一个突变体外，其他突变体都存在错定位，以及核定位。通过反激活实验，所有EBF3突变体都表现出显著降低或不具有激活报告基因CDKN1A转录的能力，原位亚细胞分离实验表明，EBF3突变体蛋白与染色质的相关性较低。最后，在RNA-seq和ChIP-seq实验中，EBF3作为转录调控因子，突变体EBF3全基因组DNA结合和基因调控活性降低。我们的研究结果表明，破坏ebf3介导的转录调节的变异会导致智力残疾和发育迟缓，并且存在于约0.1%的不明原因神经发育障碍患者中。
From a GeneMatcher-enabled international collaboration, we identified ten individuals affected by intellectual disability, speech delay, ataxia, and facial dysmorphism and carrying a deleterious EBF3 variant detected by whole-exome sequencing. One 9-bp duplication and one splice-site, five missense, and two nonsense variants in EBF3 were found; the mutations occurred de novo in eight individuals, and the missense variant c.625C>T (p.Arg209Trp) was inherited by two affected siblings from their healthy mother, who is mosaic. EBF3 belongs to the early B cell factor family (also known as Olf, COE, or O/E) and is a transcription factor involved in neuronal differentiation and maturation. Structural assessment predicted that the five amino acid substitutions have damaging effects on DNA binding of EBF3. Transient expression of EBF3 mutant proteins in HEK293T cells revealed mislocalization of all but one mutant in the cytoplasm, as well as nuclear localization. By transactivation assays, all EBF3 mutants showed significantly reduced or no ability to activate transcription of the reporter gene CDKN1A, and in situ subcellular fractionation experiments demonstrated that EBF3 mutant proteins were less tightly associated with chromatin. Finally, in RNA-seq and ChIP-seq experiments, EBF3 acted as a transcriptional regulator, and mutant EBF3 had reduced genome-wide DNA binding and gene-regulatory activity. Our findings demonstrate that variants disrupting EBF3-mediated transcriptional regulation cause intellectual disability and developmental delay and are present in ~0.1% of individuals with unexplained neurodevelopmental disorders.