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Defects in mTR stability and telomerase activity produced by the Dkc1 A353V mutation in dyskeratosis congenita are rescued by a peptide from the dyskerin TruB domain
- Source :
- Digital.CSIC. Repositorio Institucional del CSIC, instname
- Publication Year :
- 2012
- Publisher :
- Springer Science and Business Media LLC, 2012.
-
Abstract
- [Background]: The predominant X-linked form of dyskeratosis congenita results from mutations in dyskerin, a protein required for ribosomal RNA modification that is also a component of the telomerase complex. We have previously found that expression of an internal fragment of dyskerin (GSE24.2) rescues telomerase activity in X-linked dyskeratosis congenita (X-DC) patient cells. [Materials and Methods]: Here, we have generated F9 mouse cell lines expressing the most frequent mutation found in X-DC patients, A353V and study the effect of expressing the GSE24.2 cDNA or GSE24.2 peptide on telomerase activity by TRAP assay, and mTERT and mTR expression by Q-PCR. Point mutation in GSE24.2 residues were generated by site-directed mutagenesis. [Results]: Expression of GSE24.2 increases mTR and to a lesser extent mTERT RNA levels, and leads to recovery of telomerase activity. Point mutations in GSE24.2 residues known to be highly conserved and crucial for the pseudouridine-synthase activity of dyskerin abolished the effect of the peptide. Recovery of telomerase activity and increase in mTERT levels were found when the GSE24.2 peptide purified from bacteria was introduced into the cells. Moreover, mTR stability was also rescued by transfection of the peptide GSE24.2. [Discussion]: These data indicate that supplying GSE24.2, either from a cDNA vector, or as a peptide, can reduces the pathogenic effects of Dkc1 mutations and could form the basis of a novel therapeutic approach.<br />This work was supported by grants: 08/1485 from FIS, Fundación Genoma and BFU-05-0138 and Fundación Ramón Areces and R01 CA 106995 from the NIH. R.M-P was supported by Fundación Genoma. C. Manguan-Garcia and J. Carrillo were supported by CIBER de Enfermedades Raras.
- Subjects :
- Cancer Research
Telomerase
RNA Stability
Molecular Sequence Data
Mutation, Missense
Mutagenesis (molecular biology technique)
Cell Cycle Proteins
Biology
medicine.disease_cause
Dyskeratosis Congenita
Article
Dyskerin
Mice
medicine
Animals
Humans
Amino Acid Sequence
Intramolecular Transferases
Peptide sequence
Cells, Cultured
Mutation
Alanine
Point mutation
Nuclear Proteins
RNA
Valine
Genetic Therapy
General Medicine
medicine.disease
Molecular biology
Peptide Fragments
Protein Structure, Tertiary
Enzyme Activation
Amino Acid Substitution
Oncology
Dyskeratosis congenita
HeLa Cells
Subjects
Details
- ISSN :
- 16993055 and 1699048X
- Volume :
- 14
- Database :
- OpenAIRE
- Journal :
- Clinical and Translational Oncology
- Accession number :
- edsair.doi.dedup.....562ca7bd646ef3076c296f3492e9f2d5
- Full Text :
- https://doi.org/10.1007/s12094-012-0865-4