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hnRNPA2/B1 and nELAV proteins bind to a specific U-rich element in CDK5R1 3'-UTR and oppositely regulate its expression.
Biochim Biophys Acta. 2014 Jun; 1839(6):506-16.BB

Abstract

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.

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Publisher Full Text (DOI)

Authors+Show Affiliations

Zuccotti P
Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, Università degli Studi di Milano, Milan, Italy.
Colombrita C
Dipartimento di Fisiopatologia Medico-Chirurgica e dei Trapianti, Università degli Studi di Milano, Milan, Italy; IRCCS Istituto Auxologico Italiano, Cusano, Milan, Italy.
Moncini S
Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, Università degli Studi di Milano, Milan, Italy.
Barbieri A
Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, Università degli Studi di Milano, Milan, Italy.
Lunghi M
Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, Università degli Studi di Milano, Milan, Italy.
Gelfi C
Dipartimento di Scienze Biomediche per la Salute, Università degli Studi di Milano, Milan, Italy; IBFM-CNR, Segrate, Milan, Italy.
De Palma S
Dipartimento di Scienze Biomediche per la Salute, Università degli Studi di Milano, Milan, Italy; IBFM-CNR, Segrate, Milan, Italy.
Nicolin A
Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, Università degli Studi di Milano, Milan, Italy.
Ratti A
Dipartimento di Fisiopatologia Medico-Chirurgica e dei Trapianti, Università degli Studi di Milano, Milan, Italy; IRCCS Istituto Auxologico Italiano, Cusano, Milan, Italy.
Venturin M
Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, Università degli Studi di Milano, Milan, Italy. Electronic address: marco.venturin@unimi.it.
Riva P
Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, Università degli Studi di Milano, Milan, Italy. Electronic address: paola.riva@unimi.it.

MeSH

3' Untranslated RegionsBlotting, WesternCell DifferentiationELAV ProteinsGene Expression Regulation, NeoplasticHeterogeneous-Nuclear Ribonucleoprotein Group A-BHumansImmunoprecipitationLuciferasesNerve Tissue ProteinsNeuroblastomaRNA StabilityRNA, MessengerReal-Time Polymerase Chain ReactionRegulatory Sequences, Ribonucleic AcidReverse Transcriptase Polymerase Chain ReactionSpectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationTumor Cells, Cultured

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

24792867

Citation

Zuccotti, Paola, et al. "HnRNPA2/B1 and nELAV Proteins Bind to a Specific U-rich Element in CDK5R1 3'-UTR and Oppositely Regulate Its Expression." Biochimica Et Biophysica Acta, vol. 1839, no. 6, 2014, pp. 506-16.
Zuccotti P, Colombrita C, Moncini S, et al. HnRNPA2/B1 and nELAV proteins bind to a specific U-rich element in CDK5R1 3'-UTR and oppositely regulate its expression. Biochim Biophys Acta. 2014;1839(6):506-16.
Zuccotti, P., Colombrita, C., Moncini, S., Barbieri, A., Lunghi, M., Gelfi, C., De Palma, S., Nicolin, A., Ratti, A., Venturin, M., & Riva, P. (2014). HnRNPA2/B1 and nELAV proteins bind to a specific U-rich element in CDK5R1 3'-UTR and oppositely regulate its expression. Biochimica Et Biophysica Acta, 1839(6), 506-16. https://doi.org/10.1016/j.bbagrm.2014.04.018
Zuccotti P, et al. HnRNPA2/B1 and nELAV Proteins Bind to a Specific U-rich Element in CDK5R1 3'-UTR and Oppositely Regulate Its Expression. Biochim Biophys Acta. 2014;1839(6):506-16. PubMed PMID: 24792867.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - hnRNPA2/B1 and nELAV proteins bind to a specific U-rich element in CDK5R1 3'-UTR and oppositely regulate its expression. AU - Zuccotti,Paola, AU - Colombrita,Claudia, AU - Moncini,Silvia, AU - Barbieri,Andrea, AU - Lunghi,Marta, AU - Gelfi,Cecilia, AU - De Palma,Sara, AU - Nicolin,Angelo, AU - Ratti,Antonia, AU - Venturin,Marco, AU - Riva,Paola, Y1 - 2014/05/02/ PY - 2013/11/08/received PY - 2014/04/09/revised PY - 2014/04/23/accepted PY - 2014/5/6/entrez PY - 2014/5/6/pubmed PY - 2014/7/18/medline KW - CDK5R1/p35 KW - Post-transcriptional regulation KW - RNA binding protein KW - U-rich element KW - hnRNPA2/B1 KW - nELAV SP - 506 EP - 16 JF - Biochimica et biophysica acta JO - Biochim Biophys Acta VL - 1839 IS - 6 N2 - 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. SN - 0006-3002 UR - https://www.unboundmedicine.com/medline/citation/24792867/hnRNPA2/B1_and_nELAV_proteins_bind_to_a_specific_U_rich_element_in_CDK5R1_3'_UTR_and_oppositely_regulate_its_expression_ DB - PRIME DP - Unbound Medicine ER -