We have located links that may give you full text access.
Effect of c.1431C>T mutation, a causative mutation of Glanzmann's thrombasthenia, on ITGB3 splicing, gene and protein expression.
Gene 2023 September 15
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 293T cells using qPCR and Western blotting.
RESULTS: Minigene splicing assay showed that c.1431C>T mutation causes three kinds of alternative splicing;(1) a 95bp deletion in the middle of exon10, (2) a 155bp deletion (95bp deletion in the middle of exon10 plus a 60bp deletion in the right side of exon10), and (3) a 261bp 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.
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 293T cells using qPCR and Western blotting.
RESULTS: Minigene splicing assay showed that c.1431C>T mutation causes three kinds of alternative splicing;(1) a 95bp deletion in the middle of exon10, (2) a 155bp deletion (95bp deletion in the middle of exon10 plus a 60bp deletion in the right side of exon10), and (3) a 261bp 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.
Full text links
Related Resources
Trending Papers
Advances in Clinical Cardiology 2023: A Summary of Key Clinical Trials.Advances in Therapy 2024 May 15
Nutrition in the intensive care unit: from the acute phase to beyond.Intensive Care Medicine 2024 May 22
The Therapy and Management of Heart Failure with Preserved Ejection Fraction: New Insights on Treatment.Cardiac Failure Review 2024
Sodium-glucose co-transporter protein 2 (SGLT2) inhibitors for people with chronic kidney disease and diabetes.Cochrane Database of Systematic Reviews 2024 May 22
Bronchiectasis management in adults: state of the art and future directions.European Respiratory Journal 2024 May 24
Drug Therapy for Acute and Chronic Heart Failure with Preserved Ejection Fraction with Hypertension: A State-of-the-Art Review.American Journal of Cardiovascular Drugs : Drugs, Devices, and Other Interventions 2024 April 5
Pathophysiological Link and Treatment Implication of Heart Failure and Preserved Ejection Fraction in Patients with Chronic Kidney Disease.Biomedicines 2024 April 31
Get seemless 1-tap access through your institution/university
For the best experience, use the Read mobile app
All material on this website is protected by copyright, Copyright © 1994-2024 by WebMD LLC.
This website also contains material copyrighted by 3rd parties.
By using this service, you agree to our terms of use and privacy policy.
Your Privacy Choices
You can now claim free CME credits for this literature searchClaim now
Get seemless 1-tap access through your institution/university
For the best experience, use the Read mobile app