Identification of RNA-binding proteins in exosomes capable of interacting with different types of RNA: RBP-facilitated transport of RNAs into exosomes

Autor(es):Mostrar menos -

Statello, Luisa ^[1] ; Maugeri, Marco ^[1] ; Garre, Elena ^[1] ; Nawaz, Muhammad ^[1] ; Wahlgren, Jessica ^[1] ; Papadimitriou, Alexandros ^[1] ; Lundqvist, Christina ^[1] ; Lindfors, Lennart ^[2] ; Collen, Anna ^[3] ; Sunnerhagen, Per ^[4] ; Ragusa, Marco ^[5] ; Purrello, Michele ^[5] ; Di Pietro, Cinzia ^[5] ; Tigue, Natalie ^[6] ; Valadi, Hadi ^[1]

Afiliação do(s) autor(es)

^[1] Univ Gothenburg, Sahlgrenska Acad, Dept Rheumatol & Inflammat Res, Gothenburg - Sweden

^[2] Univ Gothenburg, Dept Chem & Mol Biol, Gothenburg - Sweden

^[3] AstraZeneca AB, Dept Pharmaceut Sci, Innovat Med, Gothenburg - Sweden

^[4] Univ Gothenburg, Lundberg Lab, Dept Chem & Mol Biol, Gothenburg - Sweden

^[5] Univ Catania, Dept Biomed & Biotechnol Sci, Sect Biol & Genet G Sichel, BioMol Genome & Complex Syst BioMed Unit BMGS, Catania - Italy

^[6] MedImmune Ltd, Dept Antibody Discovery & Prot Engn, Granta Pk, Cambridge - England

• Tipo de documento: Artigo Científico
• Fonte: PLoS One; v. 13, n. 4 APR 24 2018.
• Citações Web of Science: 20

Resumo

The RNA that is packaged into exosomes is termed as exosomal-shuttle RNA (esRNA); however, the players, which take this subset of RNA (esRNA) into exosomes, remain largely unknown. We hypothesized that RNA binding proteins (RBPs) could serve as key players in this mechanism, by making complexes with RNAs and transporting them into exosomes during the biosynthesis of exosomes. Here, we demonstrate the presence of 30 RBPs in exosomes that were shown to form RNA-RBP complexes with both cellular RNA and exosomal-RNA species. To assess the involvement of these RBPs in RNA-transfer into exosomes, the gene transcripts encoding six of the proteins identified in exosomes (HSP90AB1, XPO5, hnRNPH1, hnRNPM, hnRNPA2B1, and MVP) were silenced by siRNA and subsequent effect on esRNA was assessed. A significant reduction of total esRNA was observed by post-transcriptional silencing of MVP, compared to other RBPs. Furthermore, to confirm the binding of MVP with esRNA, a biotinylated-MVP was transiently expressed in HEK293F cells. Higher levels of esRNA were recovered from MVP that was eluted from exosomes of transfected cells, as compared to those of non-transfected cells. Our data indicate that these RBPs could end up in exosomes together with RNA molecules in the form of RNA-ribonu-cleoprotein complexes, which could be important for the transport of RNAs into exosomes and the maintenance of RNAs inside exosomes. This type of maintenance may favor the shuttling of RNAs from exosomes to recipient cells in the form of stable complexes. (AU)

• Processo FAPESP: 12/24574-3 - Perfil de citocinas na infecção pelo vírus da dengue: identificação de potenciais biomarcadores para diagnóstico e prognóstico
• Beneficiário: Muhammad Nawaz
• Modalidade de apoio: Bolsas no Brasil - Doutorado Direto