RESUMEN
Spinocerebellar Ataxia Type 1 (SCA1) is an autosomal dominant neurodegenerative disorder caused by a polyglutamine expansion in the ataxin-1 protein. Recent genetic correlational studies have implicated DNA damage repair pathways in modifying the age at onset of disease symptoms in SCA1 and Huntington's Disease, another polyglutamine expansion disease. We demonstrate that both endogenous and transfected ataxin-1 localizes to sites of DNA damage, which is impaired by polyglutamine expansion. This response is dependent on ataxia-telangiectasia mutated (ATM) kinase activity. Further, we characterize an ATM phosphorylation motif within ataxin-1 at serine 188. We show reduction of the Drosophila ATM homolog levels in a ATXN1[82Q] Drosophila model through shRNA or genetic cross ameliorates motor symptoms. These findings offer a possible explanation as to why DNA repair was implicated in SCA1 pathogenesis by past studies. The similarities between the ataxin-1 and the huntingtin responses to DNA damage provide further support for a shared pathogenic mechanism for polyglutamine expansion diseases.
Asunto(s)
Proteínas de la Ataxia Telangiectasia Mutada/genética , Ataxina-1/genética , Daño del ADN , Ataxias Espinocerebelosas/genética , Secuencia de Aminoácidos , Animales , Animales Modificados Genéticamente , Proteínas de la Ataxia Telangiectasia Mutada/metabolismo , Ataxina-1/metabolismo , Línea Celular , Modelos Animales de Enfermedad , Drosophila/genética , Drosophila/metabolismo , Células Epiteliales/citología , Células Epiteliales/metabolismo , Humanos , Microscopía Confocal , Mutación , Péptidos/genética , Homología de Secuencia de Aminoácido , Transducción de Señal/genética , Ataxias Espinocerebelosas/metabolismo , Ataxias Espinocerebelosas/patología , Repeticiones de Trinucleótidos/genéticaRESUMEN
Huntingtin is a 3144 amino acid protein defined as a scaffold protein with many intracellular locations that suggest functions in these compartments. Expansion of the CAG DNA tract in the huntingtin first exon is the cause of Huntington's disease. An important tool in understanding the biological functions of huntingtin is molecular imaging at the single-cell level by microscopy and nanoscopy. The evolution of these technologies has accelerated since the Nobel Prize in Chemistry was awarded in 2014 for super-resolution nanoscopy. We are in a new era of light imaging at the single-cell level, not just for protein location, but also for protein conformation and biochemical function. Large-scale microscopy-based screening is also being accelerated by a coincident development of machine-based learning that offers a framework for truly unbiased data acquisition and analysis at very large scales. This review will summarize the newest technologies in light, electron, and atomic force microscopy in the context of unique challenges with huntingtin cell biology and biochemistry.