Impaired NHEJ repair in amyotrophic lateral sclerosis is associated with TDP-43 mutations
| Autor: | Alison Cheong, Sonam Parakh, Sina Shadfar, Prachi Mehta, Shu Yang, Anand K. Deva, Anna Konopka, Natalie Grima, Adam K. Walker, Cyril J. Jagaraj, Emma R. Perri, Suzanne M. Cutts, Tina Robinson, Garth A. Nicholson, Hamideh Shahheydari, Reka P. Toth, Shafi Jamali, Julie D. Atkin, Audrey Ragagnin, Ian P. Blair, Toby D. M. Bell, Zuzana Horejsi, Marta Vidal, Ivan Khizhnyak, Donna R. Whelan, Jasmin Galper |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Adult Male TDP-43 mutations DNA End-Joining Repair DNA damage DNA repair Mutant Biology lcsh:Geriatrics medicine.disease_cause lcsh:RC346-429 03 medical and health sciences Cellular and Molecular Neuroscience chemistry.chemical_compound Mice 0302 clinical medicine mental disorders medicine Animals Humans Super-resolution microscopy Molecular Biology NHEJ lcsh:Neurology. Diseases of the nervous system Aged Uncategorized Motor Neurons Mutation Neurodegeneration Amyotrophic Lateral Sclerosis RNA nutritional and metabolic diseases Middle Aged medicine.disease Cell biology nervous system diseases Non-homologous end joining DNA-Binding Proteins lcsh:RC952-954.6 030104 developmental biology chemistry Female Neurology (clinical) 030217 neurology & neurosurgery DNA Research Article |
| Zdroj: | Molecular Neurodegeneration, Vol 15, Iss 1, Pp 1-28 (2020) Molecular Neurodegeneration |
| ISSN: | 1750-1326 |
| DOI: | 10.1186/s13024-020-00386-4 |
| Popis: | Background Pathological forms of TAR DNA-binding protein 43 (TDP-43) are present in motor neurons of almost all amyotrophic lateral sclerosis (ALS) patients, and mutations in TDP-43 are also present in ALS. Loss and gain of TDP-43 functions are implicated in pathogenesis, but the mechanisms are unclear. While the RNA functions of TDP-43 have been widely investigated, its DNA binding roles remain unclear. However, recent studies have implicated a role for TDP-43 in the DNA damage response. Methods We used NSC-34 motor neuron-like cells and primary cortical neurons expressing wildtype TDP-43 or TDP-43 ALS associated mutants (A315T, Q331K), in which DNA damage was induced by etoposide or H2O2 treatment. We investigated the consequences of depletion of TDP-43 on DNA repair using small interfering RNAs. Specific non homologous end joining (NHEJ) reporters (EJ5GFP and EJ2GFP) and cells lacking DNA-dependent serine/threonine protein kinase (DNA-PK) were used to investigate the role of TDP-43 in DNA repair. To investigate the recruitment of TDP-43 to sites of DNA damage we used single molecule super-resolution microscopy and a co-immunoprecipitation assay. We also investigated DNA damage in an ALS transgenic mouse model, in which TDP-43 accumulates pathologically in the cytoplasm. We also examined fibroblasts derived from ALS patients bearing the TDP-43 M337V mutation for evidence of DNA damage. Results We demonstrate that wildtype TDP-43 is recruited to sites of DNA damage where it participates in classical NHEJ DNA repair. However, ALS-associated TDP-43 mutants lose this activity, which induces DNA damage. Furthermore, DNA damage is present in mice displaying TDP-43 pathology, implying an active role in neurodegeneration. Additionally, DNA damage triggers features typical of TDP-43 pathology; cytoplasmic mis-localisation and stress granule formation. Similarly, inhibition of NHEJ induces TDP-43 mis-localisation to the cytoplasm. Conclusions This study reveals that TDP-43 functions in DNA repair, but loss of this function triggers DNA damage and is associated with key pathological features of ALS. |
| Databáze: | OpenAIRE |
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