Tags

Type your tag names separated by a space and hit enter

hnRNP H1 and intronic G runs in the splicing control of the human rpL3 gene.
Biochim Biophys Acta. 2010 May-Jun; 1799(5-6):419-28.BB

Abstract

By generating mRNA containing a premature termination codon (PTC), alternative splicing (AS) can quantitatively regulate the expression of genes that are degraded by nonsense-mediated mRNA decay (NMD). We previously demonstrated that AS-induced retention of part of intron 3 of rpL3 pre-mRNA produces an mRNA isoform that contains a PTC and is targeted for decay by NMD. We also demonstrated that overexpression of rpL3 downregulates canonical splicing and upregulates the alternative splicing of its pre-mRNA. We are currently investigating the molecular mechanism underlying rpL3 autoregulation. Here we report that the heterogeneous nuclear ribonucleoprotein (hnRNP) H1 is a transacting factor able to interact in vitro and in vivo with rpL3 and with intron 3 of the rpL3 gene. We investigated the role played by hnRNP H1 in the regulation of splicing of rpL3 pre-mRNA by manipulating its expression level. Depletion of hnRNP H1 reduced the level of the PTC-containing mRNA isoform, whereas its overexpression favored the selection of the cryptic 3' splice site of intron 3. We also identified and characterized the cis-acting regulatory elements involved in hnRNP H1-mediated regulation of splicing. RNA electromobility shift assay demonstrated that hnRNP H1 specifically recognizes and binds directly to the intron 3 region that contains seven copies of G-rich elements. Site-directed mutagenesis analysis and in vivo studies showed that the G3 and G6 elements are required for hnRNP H1-mediated regulation of rpL3 pre-mRNA splicing. We propose a working model in which rpL3 recruits hnRNP H1 and, through cooperation with other splicing factors, promotes selection of the alternative splice site.

Authors+Show Affiliations

Dipartimento di Biochimica e Biotecnologie Mediche, Università Federico II, Napoli 80131, Italy.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

20100605

Citation

Russo, Annapina, et al. "HnRNP H1 and Intronic G Runs in the Splicing Control of the Human rpL3 Gene." Biochimica Et Biophysica Acta, vol. 1799, no. 5-6, 2010, pp. 419-28.
Russo A, Siciliano G, Catillo M, et al. HnRNP H1 and intronic G runs in the splicing control of the human rpL3 gene. Biochim Biophys Acta. 2010;1799(5-6):419-28.
Russo, A., Siciliano, G., Catillo, M., Giangrande, C., Amoresano, A., Pucci, P., Pietropaolo, C., & Russo, G. (2010). HnRNP H1 and intronic G runs in the splicing control of the human rpL3 gene. Biochimica Et Biophysica Acta, 1799(5-6), 419-28. https://doi.org/10.1016/j.bbagrm.2010.01.008
Russo A, et al. HnRNP H1 and Intronic G Runs in the Splicing Control of the Human rpL3 Gene. Biochim Biophys Acta. 2010 May-Jun;1799(5-6):419-28. PubMed PMID: 20100605.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - hnRNP H1 and intronic G runs in the splicing control of the human rpL3 gene. AU - Russo,Annapina, AU - Siciliano,Gabriella, AU - Catillo,Morena, AU - Giangrande,Chiara, AU - Amoresano,Angela, AU - Pucci,Pietro, AU - Pietropaolo,Concetta, AU - Russo,Giulia, Y1 - 2010/01/25/ PY - 2009/08/07/received PY - 2010/01/15/revised PY - 2010/01/19/accepted PY - 2010/1/27/entrez PY - 2010/1/27/pubmed PY - 2010/6/23/medline SP - 419 EP - 28 JF - Biochimica et biophysica acta JO - Biochim. Biophys. Acta VL - 1799 IS - 5-6 N2 - By generating mRNA containing a premature termination codon (PTC), alternative splicing (AS) can quantitatively regulate the expression of genes that are degraded by nonsense-mediated mRNA decay (NMD). We previously demonstrated that AS-induced retention of part of intron 3 of rpL3 pre-mRNA produces an mRNA isoform that contains a PTC and is targeted for decay by NMD. We also demonstrated that overexpression of rpL3 downregulates canonical splicing and upregulates the alternative splicing of its pre-mRNA. We are currently investigating the molecular mechanism underlying rpL3 autoregulation. Here we report that the heterogeneous nuclear ribonucleoprotein (hnRNP) H1 is a transacting factor able to interact in vitro and in vivo with rpL3 and with intron 3 of the rpL3 gene. We investigated the role played by hnRNP H1 in the regulation of splicing of rpL3 pre-mRNA by manipulating its expression level. Depletion of hnRNP H1 reduced the level of the PTC-containing mRNA isoform, whereas its overexpression favored the selection of the cryptic 3' splice site of intron 3. We also identified and characterized the cis-acting regulatory elements involved in hnRNP H1-mediated regulation of splicing. RNA electromobility shift assay demonstrated that hnRNP H1 specifically recognizes and binds directly to the intron 3 region that contains seven copies of G-rich elements. Site-directed mutagenesis analysis and in vivo studies showed that the G3 and G6 elements are required for hnRNP H1-mediated regulation of rpL3 pre-mRNA splicing. We propose a working model in which rpL3 recruits hnRNP H1 and, through cooperation with other splicing factors, promotes selection of the alternative splice site. SN - 0006-3002 UR - https://www.unboundmedicine.com/medline/citation/20100605/hnRNP_H1_and_intronic_G_runs_in_the_splicing_control_of_the_human_rpL3_gene_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1874-9399(10)00019-2 DB - PRIME DP - Unbound Medicine ER -