RESUMEN
Transport of RNAs to dendrites occurs in neuronal RNA granules, which allows local synthesis of specific proteins at active synapses on demand, thereby contributing to learning and memory. To gain insight into the machinery controlling dendritic mRNA localization and translation, we established a stringent protocol to biochemically purify RNA granules from rat brain. Here, we identified a specific set of interactors for two RNA-binding proteins that are known components of neuronal RNA granules, Barentsz and Staufen2. First, neuronal RNA granules are much more heterogeneous than previously anticipated, sharing only a third of the identified proteins. Second, dendritically localized mRNAs, e.g., Arc and CaMKIIα, associate selectively with distinct RNA granules. Third, our work identifies a series of factors with known roles in RNA localization, translational control, and RNA quality control that are likely to keep localized transcripts in a translationally repressed state, often in distinct types of RNPs.
Asunto(s)
Dendritas/metabolismo , Biosíntesis de Proteínas , Transporte de ARN , ARN Mensajero/metabolismo , Animales , Proteína Quinasa Tipo 2 Dependiente de Calcio Calmodulina/genética , Proteína Quinasa Tipo 2 Dependiente de Calcio Calmodulina/metabolismo , Proteínas del Citoesqueleto/genética , Proteínas del Citoesqueleto/metabolismo , Hipocampo/citología , Hipocampo/metabolismo , Proteínas del Tejido Nervioso/genética , Proteínas del Tejido Nervioso/metabolismo , Unión Proteica , Proteínas de Unión al ARN/metabolismo , Ratas , Ribonucleoproteínas/metabolismoRESUMEN
Sox transcription factors are members of the Sry-related protein family that play multiple roles mainly during development. Sox18 has been implicated in the development of hair follicles as well as the blood and lymphatic vasculature, due to the identification of mutations that result in the ragged phenotype in mice, and in the hypotrichosis lymphedema telangiectasia syndrome in humans. Sox18 consists of an N-terminal high-mobility group DNA binding and a central transactivation domain, followed by a C-terminal region of unknown function. We show here that this C-terminal domain consists of three blocks that are highly conserved within a subgroup of the Sox family, and that the central so-called charged block comprises an additional strong transactivating domain. This activity can be pinpointed to a recently described 9aa transactivation motif that mediates the interaction with the transcriptional cofactor TAF9. These result can explain previously controversial data on the functional consequences of Sox18 mutations in mice and humans.