GTF3A突变导致CVID和HSE患者DNA结合能力受损。(A) GTF3A变异的临床表型和等位基因分离的家系谱系。(B) II:2和II:3的原代成纤维细胞基因组DNA的电泳图。(C) gnomAD中GTF3A变异的CADD评分相对于小等位基因频率(MAF)， MSC(99%置信区间)为4.921。(D)由9个C2H2 zf组成的TFIIIA结构域结构示意图，其中p.C195W和p.C219R突变以红色显示。下面的数字表示氨基酸。(E)来自指定物种的TFIIIA (ZF6和ZF7)的晶簇欧米茄序列比对，灰色显示保守的C2H2残基。上面的数字表示氨基酸。(F)预测人TFIIIA蛋白C2H2 ZFs的三维结构(WT和突变体)。错义突变(p.C195W和p.C219R)用红色表示。 (G) RT-qPCR analysis of GTF3A expression in fibroblasts from II:2 and II:3 compared with HCs. (H) Endogenous TFIIIA protein expression in fibroblasts from HCs, II:2, II:3, and I:2, assessed by IB. (I) TFIIIA binding to 5S rDNA ICR assessed by EMSA on WCLs of HEK293T cells that were transfected for 16 hours with empty vector (EV) or GFP-fused TFIIIA WT, missense mutants (C195W and C219R), or ZF7KO (control) at indicated ratios. (J) Densitometric quantification of (I). (K) Expression of WT and mutant TFIIIA in the WCLs for (I), determined by IB. (L) TFIIIA binding to 5S rDNA ICR, determined by EMSA on WCLs of HEK293T cells that were transfected for 16 hours with EV or the indicated GFP-fused TFIIIA WT and mutant constructs. Each data point represents one biological replicate from three (G) or four (J) independent experiments (mean ± SD), or data shown are representative of at least two independent experiments (H, I, K, and L) (two-tailed, unpaired Student’s t test). ns, statistically not significant. Credit:科学免疫学(2022)。DOI: 10.1126 / sciimmunol.abq4531
