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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Detalles Bibliográficos
Autores: Koehorst, Emma, Núñez-Manchón, Judit|||0000-0001-5154-1454, Ballester-Lopez, Alfonsina|||0000-0002-8922-1664, Almendrote, Míriam|||0000-0003-0212-275X, Lucente, Giuseppe|||0000-0003-1120-9136, Arbex, Andrea|||0000-0002-3741-4755, Chojnacki, Jakub|||0000-0002-4457-8096, Vázquez-Manrique, Rafael P.|||0000-0002-1594-0033, Gómez-Escribano, Ana Pilar|||0000-0003-4391-5932, Pintos-Morell, Guillem|||0000-0002-9347-2386, Coll-Cantí, Jaume|||0000-0001-7128-1186, Ramos-Fransi, Alba|||0000-0002-4114-4575, Martínez-Piñeiro, Alicia|||0000-0003-1988-606X, Suelves Esteban, Mònica|||0000-0001-7095-5804, Nogales, Gisela|||0000-0002-7414-212X
Tipo de recurso: artículo
Fecha de publicación:2021
País:España
Institución:Universitat Autònoma de Barcelona
Repositorio:Dipòsit Digital de Documents de la UAB
Idioma:inglés
OAI Identifier:oai:ddd.uab.cat:282874
Acceso en línea:https://ddd.uab.cat/record/282874
https://dx.doi.org/urn:doi:10.3390/jcm10235520
Access Level:acceso abierto
Palabra clave:Antisense transcription
Myotonic dystrophies
Phenotypic modulators
Primary cell cultures
RAN translation
Descripción
Sumario: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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