Min Guo, Fangfang Li, Lingyan Ren, Guiqin You, Ying Zhang, Juan Liu, Shengwen Huang
{"title":"不孕妇女TUBB8基因c.1039A >g错义突变的功能分析。","authors":"Min Guo, Fangfang Li, Lingyan Ren, Guiqin You, Ying Zhang, Juan Liu, Shengwen Huang","doi":"10.1007/s10528-025-11152-w","DOIUrl":null,"url":null,"abstract":"<p><p>The TUBB8 gene is highly conserved in primates, and pathogenic mutations in this gene have been linked to defects in oocyte maturation, leading to infertility in women. This study aimed to identify a mutation in the TUBB8 gene in a family with female infertility and functionally validate the identified mutation to confirm its pathogenicity. Genomic DNA was extracted from the proband's peripheral blood for whole exome sequencing. DNA from the proband's parents was obtained for Sanger sequencing to trace the origin of the proband's mutation. Bioinformatics analysis, conservation analysis, and three-dimensional protein structure prediction were performed on the sequencing results. Wild-type and mutant TUBB8 expression plasmids for the identified mutation sites were constructed and transfected into HEK293T and HeLa cells. Changes in protein structure and gene expression were then assessed. The analysis revealed that the proband carried the TUBB8 mutation c.1039A > G, also present in her father and aunt. This mutation was classified as a Variant of Uncertain Significance (VUS). Protein structure prediction suggested that the TUBB8 p.N347D (c.1039A > G) mutant protein had an additional hydrogen bond compared to the wild-type protein. Still, no significant structural changes were observed in the three-dimensional model. Immunofluorescence staining showed that the TUBB8 c.1039A > G mutation did not disrupt cellular microtubule structure. In vitro assays indicated that the c.1039A > G mutation decreased mRNA and protein expression levels of TUBB8. This study describes a case of female infertility associated with a newly discovered heterozygous c.1039A > G mutation in the TUBB8 gene, which may reduce TUBB8 expression. These findings contribute to the genetic understanding and diagnosis of TUBB8-related diseases.</p>","PeriodicalId":482,"journal":{"name":"Biochemical Genetics","volume":" ","pages":""},"PeriodicalIF":1.6000,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Functional Analysis of a Novel Missense Mutation c.1039A > G of TUBB8 in Infertile Women.\",\"authors\":\"Min Guo, Fangfang Li, Lingyan Ren, Guiqin You, Ying Zhang, Juan Liu, Shengwen Huang\",\"doi\":\"10.1007/s10528-025-11152-w\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>The TUBB8 gene is highly conserved in primates, and pathogenic mutations in this gene have been linked to defects in oocyte maturation, leading to infertility in women. This study aimed to identify a mutation in the TUBB8 gene in a family with female infertility and functionally validate the identified mutation to confirm its pathogenicity. Genomic DNA was extracted from the proband's peripheral blood for whole exome sequencing. DNA from the proband's parents was obtained for Sanger sequencing to trace the origin of the proband's mutation. Bioinformatics analysis, conservation analysis, and three-dimensional protein structure prediction were performed on the sequencing results. Wild-type and mutant TUBB8 expression plasmids for the identified mutation sites were constructed and transfected into HEK293T and HeLa cells. Changes in protein structure and gene expression were then assessed. The analysis revealed that the proband carried the TUBB8 mutation c.1039A > G, also present in her father and aunt. This mutation was classified as a Variant of Uncertain Significance (VUS). Protein structure prediction suggested that the TUBB8 p.N347D (c.1039A > G) mutant protein had an additional hydrogen bond compared to the wild-type protein. Still, no significant structural changes were observed in the three-dimensional model. Immunofluorescence staining showed that the TUBB8 c.1039A > G mutation did not disrupt cellular microtubule structure. In vitro assays indicated that the c.1039A > G mutation decreased mRNA and protein expression levels of TUBB8. This study describes a case of female infertility associated with a newly discovered heterozygous c.1039A > G mutation in the TUBB8 gene, which may reduce TUBB8 expression. 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Functional Analysis of a Novel Missense Mutation c.1039A > G of TUBB8 in Infertile Women.
The TUBB8 gene is highly conserved in primates, and pathogenic mutations in this gene have been linked to defects in oocyte maturation, leading to infertility in women. This study aimed to identify a mutation in the TUBB8 gene in a family with female infertility and functionally validate the identified mutation to confirm its pathogenicity. Genomic DNA was extracted from the proband's peripheral blood for whole exome sequencing. DNA from the proband's parents was obtained for Sanger sequencing to trace the origin of the proband's mutation. Bioinformatics analysis, conservation analysis, and three-dimensional protein structure prediction were performed on the sequencing results. Wild-type and mutant TUBB8 expression plasmids for the identified mutation sites were constructed and transfected into HEK293T and HeLa cells. Changes in protein structure and gene expression were then assessed. The analysis revealed that the proband carried the TUBB8 mutation c.1039A > G, also present in her father and aunt. This mutation was classified as a Variant of Uncertain Significance (VUS). Protein structure prediction suggested that the TUBB8 p.N347D (c.1039A > G) mutant protein had an additional hydrogen bond compared to the wild-type protein. Still, no significant structural changes were observed in the three-dimensional model. Immunofluorescence staining showed that the TUBB8 c.1039A > G mutation did not disrupt cellular microtubule structure. In vitro assays indicated that the c.1039A > G mutation decreased mRNA and protein expression levels of TUBB8. This study describes a case of female infertility associated with a newly discovered heterozygous c.1039A > G mutation in the TUBB8 gene, which may reduce TUBB8 expression. These findings contribute to the genetic understanding and diagnosis of TUBB8-related diseases.
期刊介绍:
Biochemical Genetics welcomes original manuscripts that address and test clear scientific hypotheses, are directed to a broad scientific audience, and clearly contribute to the advancement of the field through the use of sound sampling or experimental design, reliable analytical methodologies and robust statistical analyses.
Although studies focusing on particular regions and target organisms are welcome, it is not the journal’s goal to publish essentially descriptive studies that provide results with narrow applicability, or are based on very small samples or pseudoreplication.
Rather, Biochemical Genetics welcomes review articles that go beyond summarizing previous publications and create added value through the systematic analysis and critique of the current state of knowledge or by conducting meta-analyses.
Methodological articles are also within the scope of Biological Genetics, particularly when new laboratory techniques or computational approaches are fully described and thoroughly compared with the existing benchmark methods.
Biochemical Genetics welcomes articles on the following topics: Genomics; Proteomics; Population genetics; Phylogenetics; Metagenomics; Microbial genetics; Genetics and evolution of wild and cultivated plants; Animal genetics and evolution; Human genetics and evolution; Genetic disorders; Genetic markers of diseases; Gene technology and therapy; Experimental and analytical methods; Statistical and computational methods.