Your browser doesn't support javascript.
loading
Research on screening and identification of proteins interacting with ataxin-3 / 中华医学遗传学杂志
Chinese Journal of Medical Genetics ; (6): 242-247, 2005.
Artículo en Inglés | WPRIM | ID: wpr-321116
ABSTRACT
<p><b>OBJECTIVE</b>This study sought to isolate and identify the proteins that interact with ataxin-3, to confirm the interacted domain, and to provide new clues for exploring the function of ataxin-3 and the pathogenesis of spinocerebellar ataxia type 3 and Machado-Joseph disease (SCA3/MJD).</p><p><b>METHODS</b>Yeast two-hybrid screen (MATCHMAKER GAL4 Two-Hybrid System 3) and regular molecular biologic techniques were undertaken to screen human brain cDNA library with mutant ataxin-3 bait. Two baits from both normal and mutant C-terminus of ataxin-3 were created by subcloned methods to determine which domain of ataxin-3 interacts with the putative associated proteins and to find out optimal candidate proteins that interact with C-terminus of ataxin-3. Confocal microscope was used to observe whether ataxin-3 co-localized with the obtained interacting proteins in mammalian cells.</p><p><b>RESULTS</b>Five novel ataxin-3 interacting proteins were obtained, among which were three known proteins, namely human rhodopsin guanosine diphosphate dissociation inhibitor alpha, small ubiquitin-like modifier 1, and human neuronal amiloride-sensitive cation channel 2; the other two were unknown. Interacting domain analysis revealed that an unknown protein interacted with the C-terminus near the polyglutamine tract of ataxin-3, the other four all interacted with the N-terminus. In the nucleus of SH-SY5Y cell, small ubiquitin-like modifier 1 co-localized with the wild-type ataxin-3 and with the intranuclear aggregates formed by the mutant ataxin-3.</p><p><b>CONCLUSION</b>An unknown protein probably interacting with C-terminus of ataxin-3 is firstly discovered, and the initiative findings suggest first that the interaction of small ubiquitin-like modifier 1 with N-terminus of ataxin-3 and the relevant sumoylation probably participate in the post-translation modifying of ataxin-3 and in the pathogenesis of SCA3/MJD.</p>
Asunto(s)
Texto completo: Disponible Índice: WPRIM (Pacífico Occidental) Asunto principal: Plásmidos / Unión Proteica / Proteínas Represoras / Proteínas Recombinantes de Fusión / Proteínas Nucleares / Transfección / Canales de Sodio / Microscopía Confocal / Técnicas del Sistema de Dos Híbridos / Proteína SUMO-1 Tipo de estudio: Estudio diagnóstico / Estudio pronóstico / Estudio de tamizaje Límite: Humanos Idioma: Inglés Revista: Chinese Journal of Medical Genetics Año: 2005 Tipo del documento: Artículo

Similares

MEDLINE

...
LILACS

LIS

Texto completo: Disponible Índice: WPRIM (Pacífico Occidental) Asunto principal: Plásmidos / Unión Proteica / Proteínas Represoras / Proteínas Recombinantes de Fusión / Proteínas Nucleares / Transfección / Canales de Sodio / Microscopía Confocal / Técnicas del Sistema de Dos Híbridos / Proteína SUMO-1 Tipo de estudio: Estudio diagnóstico / Estudio pronóstico / Estudio de tamizaje Límite: Humanos Idioma: Inglés Revista: Chinese Journal of Medical Genetics Año: 2005 Tipo del documento: Artículo