Cyclin-dependent kinase 5 regulatory subunit 1 (CDK5R1) encodes p35, a specific activator of cyclin-dependent kinase 5 (CDK5). CDK5 and p35 have a fundamental role in neuronal migration and differentiation during CNS development. Both the CDK5R1 3′-UTR's remarkable size and its conservation during evolution strongly indicate an important role in post-transcriptional regulation. We previously validated different regulatory elements in the 3′-UTR of CDK5R1, which affect transcript stability, p35 levels and cellular migration through the binding with nELAV proteins and miR-103/7 miRNAs. Interestingly, a 138 bp-long region, named C2.1, was identified as the most mRNA destabilizing portion within CDK5R1 3′-UTR. This feature was maintained by a shorter region of 73 bp, characterized by two poly-U stretches. UV-CL experiments showed that this region interacts with protein factors. UV-CLIP assays and pull-down experiments followed by mass spectrometry analysis demonstrated that nELAV and hnRNPA2/B1 proteins bind to the same U-rich element. These RNA-binding proteins (RBPs) were shown to oppositely control CDK5R1 mRNA stability and p35 protein content at post-trascriptional level. While nELAV proteins have a positive regulatory effect, hnRNPA2/B1 has a negative action that is responsible for the mRNA destabilizing activity both of the C2.1 region and of the full-length 3′-UTR. In co-expression experiments of hnRNPA2/B1 and nELAV RBPs we observed an overall decrease of p35 content. We also demonstrated that hnRNPA2/B1 can downregulate nELAV protein content but not vice versa. This study, by providing new insights on the combined action of different regulatory factors, contributes to clarify the complex post-transcriptional control of CDK5R1 gene expression.
hnRNPA2/B1 and nELAV proteins bind to a specific U-rich element in CDK5R1 3'-UTR and oppositely regulate its expression / P. Zuccotti, C. Colombrita, S. Moncini, A. Barbieri, M. Lunghi, C. Gelfi, S. De Palma, A. Nicolin, A. Ratti, M. Venturin, P. Riva. - In: BIOCHIMICA ET BIOPHYSICA ACTA. - ISSN 0006-3002. - 1839:6(2014 May 02), pp. 506-516.
|Titolo:||hnRNPA2/B1 and nELAV proteins bind to a specific U-rich element in CDK5R1 3'-UTR and oppositely regulate its expression|
COLOMBRITA, CLAUDIA (Secondo)
DE PALMA, SARA (Primo)
RATTI, ANTONIA (Corresponding)
|Parole Chiave:||CDK5R1/p35; HnRNPA2/B1; NELAV; Post-transcriptional regulation; RNA binding protein; U-rich element|
|Settore Scientifico Disciplinare:||Settore BIO/13 - Biologia Applicata|
Settore BIO/10 - Biochimica
|Data di pubblicazione:||2-mag-2014|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1016/j.bbagrm.2014.04.018|
|Appare nelle tipologie:||01 - Articolo su periodico|