The readthrough of premature termination codon (PTC) by ribosome sometimes produces full-length proteins. We previously reported a readthrough of PTC of glycosyltransferase gene B4GALNT1 with hereditary spastic paraplegia (HSP). Here we featured the readthrough of B4GALNT1 of two mutants, M4 and M2 with PTC by immunoblotting and flow cytometry after transfection of B4GALNT1 cDNAs into cells. Immunoblotting showed a faint band of full-length mutant protein of M4 but not M2 at a similar position with that of wild-type B4GALNT1. AGC sequences at immediately before and after the PTC in M4 were critical for the readthrough. Treatment of cells transfected with mutant M4 cDNA with aminoglycosides resulted in increased readthrough of PTC. Furthermore, treatment of transfectants of mutant M2 cDNA with G418 also resulted in the induction of readthrough of PTC. Both M4 and M2 cDNA transfectants showed increased/induced bands in immunoblotting and GM2 expression in a dose-dependent manner of aminoglycosides. Results of mass spectrometry supported this effect. Here, we showed for the first time the induction and/or enhancement of the readthrough of PTCs of B4GALNT1 by aminoglycoside treatment, suggesting that aminoglycosides are efficient for patients with HSP caused by PTC of B4GALNT1, in which gradual neurological disorders emerged with aging.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1093/jb/mvaa041 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Mechanism-based approach in designing patient-specific combination therapies for nonsense mutation diseases.
bioRxiv
December 2024
Department of Chemistry, University of Pennsylvania, Philadelphia, PA 19104, USA.
Premature termination codon (PTC) diseases, arising as a consequence of nonsense mutations in a patient's DNA, account for approximately 12% of all human disease mutations. Currently there are no FDA approved treatments for increasing PTC readthrough in nonsense mutation diseases, although one translational readthrough inducing drug, ataluren, has had conditional approval for treatment of Duchenne muscular dystrophy in Europe and elsewhere for 10 years. Ataluren displays consistent low toxicity in clinical trials for treatment of several different PTC diseases, but its therapeutic effects on such diseases are inconsistent.
View Article and Find Full Text PDFMisincorporations of amino acids in p53 in human cells at artificially constructed termination codons in the presence of the aminoglycoside Gentamicin.
Front Genet
November 2024
International Centre for Cancer Vaccine Science (ICCVS), University of Gdańsk, Gdańsk, Poland.
Readthrough of a translation termination codon is regulated by ribosomal A site recognition and insertion of near-cognate tRNAs. Small molecules exist that mediate incorporation of amino acids at the stop codon and production of full-length, often functional protein but defining the actual amino acid that is incorporated remains a challenging area. Herein, we report on the development a human cell model that can be used to determine whether rules can be developed using mass spectrometry that define the type of amino acid that is placed at a premature termination codon (PTC) during readthrough mediated by an aminoglycoside.
View Article and Find Full Text PDFPromoting readthrough of nonsense mutations in CF mouse model: Biodistribution and efficacy of NV848 in rescuing CFTR protein expression.
Mol Ther
December 2024
Department of Biological, Chemical and Pharmaceutical Sciences and Technologies, University of Palermo, Palermo, Italy. Electronic address:
Article Synopsis
- Nonsense mutations lead to a stop signal in protein production, resulting in non-functional proteins and contributing to diseases like cystic fibrosis (CF).
- This research explores the potential of NV848, a drug designed to promote the continuation of protein synthesis, to enhance CFTR protein expression in a mouse model with a specific CF-causing mutation.
- The study evaluates the drug's stability, distribution in the body, and effectiveness in restoring CFTR protein in mice, suggesting NV848 could be a valuable treatment for CF patients with nonsense mutations.
Transition and management of patients with Duchenne Muscular Dystrophy: a narrative review based on Italian experts' opinion and real-world experience.
Acta Myol
September 2024
Child Neurology Unit, Presidio Ospedaliero Provinciale Santa Maria Nuova, AUSL-IRCCS di Reggio Emilia, Reggio Emilia, Italy.
Article Synopsis
- Duchenne Muscular Dystrophy (DMD) is a serious, progressive disorder that leads to muscle wasting and other complications, requiring a combination of therapies and new approaches to patient care.
- Experts in Italy discussed the challenges of transitioning care for DMD patients from pediatric to adult services, emphasizing the importance of continuous treatment and tracking relevant health outcomes after patients lose their ability to walk.
- Following loss of ambulation, care shifts focus toward cardiac and respiratory health, nutrition, and the maintenance of upper limb function, highlighting the need for shared protocols and better data collection for optimized management.
Novel readthrough agent suppresses nonsense mutations and restores functional type VII collagen and laminin 332 in epidermolysis bullosa.
Mol Ther Nucleic Acids
December 2024
Department of Dermatology, The Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA.
Recessive dystrophic epidermolysis bullosa (RDEB) and junctional epidermolysis bullosa (JEB) are lethal blistering skin disorders resulting from mutations in genes coding for type VII collagen () and laminin 332 (, , or ), respectively. In RDEB, 25% of patients harbor nonsense mutations causing premature termination codons (PTCs). In JEB, a majority of mutations in are nonsense mutations (80%).
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!