Bo Huang , Shiwei Li , Yun Chai , Yu Fan, Xin Li, Yue Liu, Yunhong Fu, Xixi Song, Jingqiu Cui
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Three-dimensional modeling, luciferase-reporter gene test, western blotting and cellular immunofluorescence were used to evaluate the effect of the mutation.</p></div><div><h3>Results</h3><p>A novel frameshift mutation c. 677dup(p.Pro227AlafsTer77), named P227Afs, was found in <em>GATA3</em>. Three-dimensional modeling revealed that the mutation caused the loss of the dual zinc finger structures 1 and 2 (ZNF1 and ZNF2) of the synthesized protein. Expression of wild-type GATA3 produced a six-fold increase in luciferase activity when compared with pcDNA3.1 vector only (<em>P</em> < 0.001), whereas the P227Afs mutant showed no increase. The mutation significantly reduced the transcriptional activity of <em>GATA3</em>. Immunofluorescence and western blotting analyses demonstrated that the mutation changed the nuclear location of GATA3 and caused difficulty in nuclearization.</p></div><div><h3>Conclusion</h3><p>A novel heterozygous frameshift mutation in <em>GATA3</em> was identified and showed to result in difficult nuclearization, and a dominant-negative effect on the wild-type.</p></div>","PeriodicalId":18814,"journal":{"name":"Molecular Genetics and Metabolism Reports","volume":null,"pages":null},"PeriodicalIF":1.8000,"publicationDate":"2024-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2214426924000168/pdfft?md5=521b2443814e15ed421259ec40ed5e67&pid=1-s2.0-S2214426924000168-main.pdf","citationCount":"0","resultStr":"{\"title\":\"A novel GATA3 frameshift mutation causes hypoparathyroidism, sensorineural deafness, and renal dysplasia syndrome\",\"authors\":\"Bo Huang , Shiwei Li , Yun Chai , Yu Fan, Xin Li, Yue Liu, Yunhong Fu, Xixi Song, Jingqiu Cui\",\"doi\":\"10.1016/j.ymgmr.2024.101063\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Background</h3><p>Hypoparathyroidism, sensorineural deafness, and renal dysplasia (HDR) syndrome (Barakat syndrome) is a rare autosomal dominant disorder caused by mutations in the gene encoding <em>GATA3</em> on chromosome 10p14.</p></div><div><h3>Method</h3><p>Informed consent was obtained from a 38-year-old female patient. 5 mL of venous blood was collected and sent for whole-exome sequencing. <em>GATA3</em> constructs of both wild-type and mutant were transfected into HEK-293 T cells. Three-dimensional modeling, luciferase-reporter gene test, western blotting and cellular immunofluorescence were used to evaluate the effect of the mutation.</p></div><div><h3>Results</h3><p>A novel frameshift mutation c. 677dup(p.Pro227AlafsTer77), named P227Afs, was found in <em>GATA3</em>. Three-dimensional modeling revealed that the mutation caused the loss of the dual zinc finger structures 1 and 2 (ZNF1 and ZNF2) of the synthesized protein. Expression of wild-type GATA3 produced a six-fold increase in luciferase activity when compared with pcDNA3.1 vector only (<em>P</em> < 0.001), whereas the P227Afs mutant showed no increase. The mutation significantly reduced the transcriptional activity of <em>GATA3</em>. Immunofluorescence and western blotting analyses demonstrated that the mutation changed the nuclear location of GATA3 and caused difficulty in nuclearization.</p></div><div><h3>Conclusion</h3><p>A novel heterozygous frameshift mutation in <em>GATA3</em> was identified and showed to result in difficult nuclearization, and a dominant-negative effect on the wild-type.</p></div>\",\"PeriodicalId\":18814,\"journal\":{\"name\":\"Molecular Genetics and Metabolism Reports\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":1.8000,\"publicationDate\":\"2024-01-30\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.sciencedirect.com/science/article/pii/S2214426924000168/pdfft?md5=521b2443814e15ed421259ec40ed5e67&pid=1-s2.0-S2214426924000168-main.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Molecular Genetics and Metabolism Reports\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2214426924000168\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"GENETICS & HEREDITY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Molecular Genetics and Metabolism Reports","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2214426924000168","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"GENETICS & HEREDITY","Score":null,"Total":0}
引用次数: 0
摘要
背景甲状旁腺功能亢进、感音神经性耳聋和肾发育不良(HDR)综合征(Barakat 综合征)是一种罕见的常染色体显性遗传疾病,由染色体 10p14 上编码 GATA3 的基因突变引起。采集 5 mL 静脉血并送去进行全外显子组测序。将野生型和突变型的 GATA3 构建体转染到 HEK-293 T 细胞中。结果 在 GATA3 中发现了一个名为 P227Afs 的新型换框突变 c. 677dup(p.Pro227AlafsTer77)。三维建模显示,该突变导致合成蛋白的双锌指结构 1 和 2(ZNF1 和 ZNF2)缺失。与仅表达 pcDNA3.1 载体相比,表达野生型 GATA3 所产生的荧光素酶活性增加了六倍(P < 0.001),而 P227Afs 突变体则没有增加。该突变明显降低了 GATA3 的转录活性。免疫荧光和 Western 印迹分析表明,该突变改变了 GATA3 的核位置,导致核化困难。
A novel GATA3 frameshift mutation causes hypoparathyroidism, sensorineural deafness, and renal dysplasia syndrome
Background
Hypoparathyroidism, sensorineural deafness, and renal dysplasia (HDR) syndrome (Barakat syndrome) is a rare autosomal dominant disorder caused by mutations in the gene encoding GATA3 on chromosome 10p14.
Method
Informed consent was obtained from a 38-year-old female patient. 5 mL of venous blood was collected and sent for whole-exome sequencing. GATA3 constructs of both wild-type and mutant were transfected into HEK-293 T cells. Three-dimensional modeling, luciferase-reporter gene test, western blotting and cellular immunofluorescence were used to evaluate the effect of the mutation.
Results
A novel frameshift mutation c. 677dup(p.Pro227AlafsTer77), named P227Afs, was found in GATA3. Three-dimensional modeling revealed that the mutation caused the loss of the dual zinc finger structures 1 and 2 (ZNF1 and ZNF2) of the synthesized protein. Expression of wild-type GATA3 produced a six-fold increase in luciferase activity when compared with pcDNA3.1 vector only (P < 0.001), whereas the P227Afs mutant showed no increase. The mutation significantly reduced the transcriptional activity of GATA3. Immunofluorescence and western blotting analyses demonstrated that the mutation changed the nuclear location of GATA3 and caused difficulty in nuclearization.
Conclusion
A novel heterozygous frameshift mutation in GATA3 was identified and showed to result in difficult nuclearization, and a dominant-negative effect on the wild-type.
期刊介绍:
Molecular Genetics and Metabolism Reports is an open access journal that publishes molecular and metabolic reports describing investigations that use the tools of biochemistry and molecular biology for studies of normal and diseased states. In addition to original research articles, sequence reports, brief communication reports and letters to the editor are considered.