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Characterization of RAN Translation and Antisense Transcription in Primary Cell Cultures of Patients with Myotonic Dystrophy Type 1

Myotonic Dystrophy type 1 (DM1) is a muscular dystrophy with a multi-systemic nature. It was one of the first diseases in which repeat associated non-ATG (RAN) translation was described in 2011, but has not been further explored since. In order to enhance our knowledge of RAN translation in DM1, we...

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Autores principales: Koehorst, Emma, Núñez-Manchón, Judit, Ballester-López, Alfonsina, Almendrote, Miriam, Lucente, Giuseppe, Arbex, Andrea, Chojnacki, Jakub, Vázquez-Manrique, Rafael P., Gómez-Escribano, Ana Pilar, Pintos-Morell, Guillem, Coll-Cantí, Jaume, Ramos-Fransi, Alba, Martínez-Piñeiro, Alicia, Suelves, Mònica, Nogales-Gadea, Gisela
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8658563/
https://www.ncbi.nlm.nih.gov/pubmed/34884222
http://dx.doi.org/10.3390/jcm10235520
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author Koehorst, Emma
Núñez-Manchón, Judit
Ballester-López, Alfonsina
Almendrote, Miriam
Lucente, Giuseppe
Arbex, Andrea
Chojnacki, Jakub
Vázquez-Manrique, Rafael P.
Gómez-Escribano, Ana Pilar
Pintos-Morell, Guillem
Coll-Cantí, Jaume
Ramos-Fransi, Alba
Martínez-Piñeiro, Alicia
Suelves, Mònica
Nogales-Gadea, Gisela
author_facet Koehorst, Emma
Núñez-Manchón, Judit
Ballester-López, Alfonsina
Almendrote, Miriam
Lucente, Giuseppe
Arbex, Andrea
Chojnacki, Jakub
Vázquez-Manrique, Rafael P.
Gómez-Escribano, Ana Pilar
Pintos-Morell, Guillem
Coll-Cantí, Jaume
Ramos-Fransi, Alba
Martínez-Piñeiro, Alicia
Suelves, Mònica
Nogales-Gadea, Gisela
author_sort Koehorst, Emma
collection PubMed
description Myotonic Dystrophy type 1 (DM1) is a muscular dystrophy with a multi-systemic nature. It was one of the first diseases in which repeat associated non-ATG (RAN) translation was described in 2011, but has not been further explored since. In order to enhance our knowledge of RAN translation in DM1, we decided to study the presence of DM1 antisense (DM1-AS) transcripts (the origin of the polyglutamine (polyGln) RAN protein) using RT-PCR and FISH, and that of RAN translation via immunoblotting and immunofluorescence in distinct DM1 primary cell cultures, e.g., myoblasts, skin fibroblasts and lymphoblastoids, from ten patients. DM1-AS transcripts were found in all DM1 cells, with a lower expression in patients compared to controls. Antisense RNA foci were found in the nuclei and cytoplasm of a subset of DM1 cells. The polyGln RAN protein was undetectable in all three cell types with both approaches. Immunoblots revealed a 42 kD polyGln containing protein, which was most likely the TATA-box-binding protein. Immunofluorescence revealed a cytoplasmic aggregate, which co-localized with the Golgi apparatus. Taken together, DM1-AS transcript levels were lower in patients compared to controls and a small portion of the transcripts included the expanded repeat. However, RAN translation was not present in patient-derived DM1 cells, or was in undetectable quantities for the available methods.
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spelling pubmed-86585632021-12-10 Characterization of RAN Translation and Antisense Transcription in Primary Cell Cultures of Patients with Myotonic Dystrophy Type 1 Koehorst, Emma Núñez-Manchón, Judit Ballester-López, Alfonsina Almendrote, Miriam Lucente, Giuseppe Arbex, Andrea Chojnacki, Jakub Vázquez-Manrique, Rafael P. Gómez-Escribano, Ana Pilar Pintos-Morell, Guillem Coll-Cantí, Jaume Ramos-Fransi, Alba Martínez-Piñeiro, Alicia Suelves, Mònica Nogales-Gadea, Gisela J Clin Med Article Myotonic Dystrophy type 1 (DM1) is a muscular dystrophy with a multi-systemic nature. It was one of the first diseases in which repeat associated non-ATG (RAN) translation was described in 2011, but has not been further explored since. In order to enhance our knowledge of RAN translation in DM1, we decided to study the presence of DM1 antisense (DM1-AS) transcripts (the origin of the polyglutamine (polyGln) RAN protein) using RT-PCR and FISH, and that of RAN translation via immunoblotting and immunofluorescence in distinct DM1 primary cell cultures, e.g., myoblasts, skin fibroblasts and lymphoblastoids, from ten patients. DM1-AS transcripts were found in all DM1 cells, with a lower expression in patients compared to controls. Antisense RNA foci were found in the nuclei and cytoplasm of a subset of DM1 cells. The polyGln RAN protein was undetectable in all three cell types with both approaches. Immunoblots revealed a 42 kD polyGln containing protein, which was most likely the TATA-box-binding protein. Immunofluorescence revealed a cytoplasmic aggregate, which co-localized with the Golgi apparatus. Taken together, DM1-AS transcript levels were lower in patients compared to controls and a small portion of the transcripts included the expanded repeat. However, RAN translation was not present in patient-derived DM1 cells, or was in undetectable quantities for the available methods. MDPI 2021-11-25 /pmc/articles/PMC8658563/ /pubmed/34884222 http://dx.doi.org/10.3390/jcm10235520 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Koehorst, Emma
Núñez-Manchón, Judit
Ballester-López, Alfonsina
Almendrote, Miriam
Lucente, Giuseppe
Arbex, Andrea
Chojnacki, Jakub
Vázquez-Manrique, Rafael P.
Gómez-Escribano, Ana Pilar
Pintos-Morell, Guillem
Coll-Cantí, Jaume
Ramos-Fransi, Alba
Martínez-Piñeiro, Alicia
Suelves, Mònica
Nogales-Gadea, Gisela
Characterization of RAN Translation and Antisense Transcription in Primary Cell Cultures of Patients with Myotonic Dystrophy Type 1
title Characterization of RAN Translation and Antisense Transcription in Primary Cell Cultures of Patients with Myotonic Dystrophy Type 1
title_full Characterization of RAN Translation and Antisense Transcription in Primary Cell Cultures of Patients with Myotonic Dystrophy Type 1
title_fullStr Characterization of RAN Translation and Antisense Transcription in Primary Cell Cultures of Patients with Myotonic Dystrophy Type 1
title_full_unstemmed Characterization of RAN Translation and Antisense Transcription in Primary Cell Cultures of Patients with Myotonic Dystrophy Type 1
title_short Characterization of RAN Translation and Antisense Transcription in Primary Cell Cultures of Patients with Myotonic Dystrophy Type 1
title_sort characterization of ran translation and antisense transcription in primary cell cultures of patients with myotonic dystrophy type 1
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8658563/
https://www.ncbi.nlm.nih.gov/pubmed/34884222
http://dx.doi.org/10.3390/jcm10235520
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