دورية أكاديمية
Effect of c.1431C > T mutation, a causative mutation of Glanzmann's thrombasthenia, on ITGB3 splicing, gene and protein expression.
| العنوان: | Effect of c.1431C > T mutation, a causative mutation of Glanzmann's thrombasthenia, on ITGB3 splicing, gene and protein expression. |
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| المؤلفون: | Wang D; Department of Pediatrics, Jinhua Maternal and Child Health Care Hospital, Jinhua Hospital of Zhejiang University School of Medicine, Jinhua City 321000, Zhejiang Province, China., Lai P; Department of Pediatrics, Jinhua Maternal and Child Health Care Hospital, Jinhua Hospital of Zhejiang University School of Medicine, Jinhua City 321000, Zhejiang Province, China., Lu Q; Department of Pediatrics, Jinhua Maternal and Child Health Care Hospital, Jinhua Hospital of Zhejiang University School of Medicine, Jinhua City 321000, Zhejiang Province, China., El-Magd MA; Department of Anatomy & Embryology, Faculty of Veterinary Medicine, Kafrelsheikh University, Egypt. Electronic address: mohamed.abouelmagd@vet.kfs.edu.eg., Li X; Department of Pediatrics, Jinhua Maternal and Child Health Care Hospital, Jinhua Hospital of Zhejiang University School of Medicine, Jinhua City 321000, Zhejiang Province, China. Electronic address: lixiaobing1110@163.com. |
| المصدر: | Gene [Gene] 2023 Dec 20; Vol. 888, pp. 147805. Date of Electronic Publication: 2023 Sep 15. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Elsevier/North-Holland Country of Publication: Netherlands NLM ID: 7706761 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1879-0038 (Electronic) Linking ISSN: 03781119 NLM ISO Abbreviation: Gene Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Amsterdam, Elsevier/North-Holland, 1976- |
| مواضيع طبية MeSH: | Thrombasthenia*/genetics , Thrombasthenia*/diagnosis, Humans ; Mutation ; RNA Splicing ; Base Sequence ; RNA, Messenger/genetics ; Integrin beta3/genetics |
| مستخلص: | Background/aim: Recently, it was reported that the non-synonymous c.1431C > T (p. G477=) mutation of the integrin subunit β3 (ITGB3) gene is the cause of Glanzmann's thrombasthenia (GT). However, the functional consequences of this mutation on the ITGB3 gene and protein expression remain to be elucidated. Therefore, this study was conducted to cover this scientific shortage. Methods: Peripheral blood samples were collected from Chinese family members (parents and proband and his sister), and DNA was extracted and sequenced using whole-exome and Sanger sequencing. The effect of c.1431C > T mutation on the splicing of mRNA was verified by the in vitro minigene assay and the three variants that resulted from the mutation were cloned into a phage vector and pEGFP-C1 vector, and ITGB3 gene and protein expression was detected in the transfected 293 T cells using qPCR and Western blotting. Results: Minigene splicing assay showed that c.1431C > T mutation causes three kinds of alternative splicing; (1) a 95 bp deletion in the middle of exon10, (2) a 155 bp deletion (95 bp deletion in the middle of exon10 plus a 60 bp deletion in the right side of exon10), and (3) a 261 bp deletion in the right side of exon10. The in vitro expression assay showed that the c.1431C > T variant did not affect the ITGB3 mRNA levels, but directly led to protein truncation and declined expression. Conclusion: Due to its significant impact on protein expression, c.1431C > T mutation in ITGB3 could be considered a pathogenic variant of GT. This could enrich the ITGB3 mutation spectrum and provide a base for the genetic diagnosis of GT. Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. (Copyright © 2023 Elsevier B.V. All rights reserved.) |
| فهرسة مساهمة: | Keywords: Aberrant splicing; Glanzmann’s thrombasthenia; ITGB3; Minigene; c.1431C>T mutation |
| المشرفين على المادة: | 0 (RNA, Messenger) 0 (ITGB3 protein, human) 0 (Integrin beta3) |
| تواريخ الأحداث: | Date Created: 20230916 Date Completed: 20231002 Latest Revision: 20231010 |
| رمز التحديث: | 20231215 |
| DOI: | 10.1016/j.gene.2023.147805 |
| PMID: | 37716584 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1879-0038 |
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| DOI: | 10.1016/j.gene.2023.147805 |