| Autor: |
Barbato C; Institute of Biochemistry and Cell Biology, National Research Council CNR, Department of Sense Organs, University of Rome Sapienza, 00161 Roma, Italy., Frisone P; Institute of Biochemistry and Cell Biology CNR, Campus A. Buzzati-Traverso, 00015 Monterotondo (RM), Italy.; Department of Molecular Medicine, University of Rome Sapienza, 00161 Roma, Italy., Braccini L; Department of Molecular Medicine, University of Rome Sapienza, 00161 Roma, Italy., D'Aguanno S; Preclinical Models and New Therapeutic Agents Unit, IRCCS Regina Elena National Cancer Institute, 00144 Rome, Italy., Pieroni L; Department of Experimental Neuroscience, Proteomics and Metabolomics Unit, IRCCS-Fondazione Santa Lucia, 00143 Rome, Italy., Ciotti MT; Institute of Biochemistry and Cell Biology CNR, Campus A. Buzzati-Traverso, 00015 Monterotondo (RM), Italy., Catalanotto C; Department of Molecular Medicine, University of Rome Sapienza, 00161 Roma, Italy., Cogoni C; Department of Molecular Medicine, University of Rome Sapienza, 00161 Roma, Italy., Ruberti F; Institute of Biochemistry and Cell Biology CNR, Campus A. Buzzati-Traverso, 00015 Monterotondo (RM), Italy. |
| Abstrakt: |
RNA-binding proteins (RBPs) play important roles in modulating miRNA-mediated mRNA target repression. Argonaute2 (Ago2) is an essential component of the RNA-induced silencing complex (RISC) that plays a central role in silencing mechanisms via small non-coding RNA molecules known as siRNAs and miRNAs. Small RNAs loaded into Argonaute proteins catalyze endoribonucleolytic cleavage of target RNAs or recruit factors responsible for translational silencing and mRNA target destabilization. In previous studies we have shown that KCC2, a neuronal Cl (-) extruding K (+) Cl (-) co-transporter 2, is regulated by miR-92 in neuronal cells. Searching for Ago2 partners by immunoprecipitation and LC-MS/MS analysis, we isolated among other proteins the Serpine mRNA binding protein 1 (SERBP1) from SH-SY5Y neuroblastoma cells. Exploring the role of SERBP1 in miRNA-mediated gene silencing in SH-SY5Y cells and primary hippocampal neurons, we demonstrated that SERBP1 silencing regulates KCC2 expression through the 3' untranslated region (UTR). In addition, we found that SERBP1 as well as Ago2/miR-92 complex bind to KCC2 3'UTR. Finally, we demonstrated the attenuation of miR-92-mediated repression of KCC2 3'UTR by SERBP1 silencing. These findings advance our knowledge regarding the miR-92-mediated modulation of KCC2 translation in neuronal cells and highlight SERBP1 as a key component of this gene regulation. |
