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hnRNP I/PTB can antagonize the splicing repressor activity of SRp30c.
RNA. 2007 Aug; 13(8):1287-300.RNA

Abstract

The control of alternative pre-mRNA splicing often requires the participation of factors displaying synergistic or antagonistic activities. In the hnRNP A1 pre-mRNA, three elements promote the exclusion of alternative exon 7B, while a fourth intron element (CE9) represses splicing of exon 7B to the downstream exon. We have shown previously that the 5' portion of the 38-nucleotide-long CE9 element is bound by SRp30c, and that this interaction is important for repression in vitro. To determine whether SRp30c alone can impose repression, we tested a high-affinity SRp30c binding site that we identified using the SELEX protocol. We find that multiple high-affinity SRp30c sites are required to replicate the level of repression obtained with CE9, and that both the 5' and the 3' portions of CE9 contribute to SRp30c binding. Performing RNA affinity chromatography with the complete CE9 element recovered hnRNP I/PTB. Surprisingly however, His-tagged PTB reduced the binding of SRp30c to CE9 in a nuclear extract, stimulated splicing to a downstream 3' splice site, and relieved the CE9-mediated splicing repression in vitro. Our in vivo results are consistent with the notion that increasing PTB levels alleviates the repression imposed by CE9 to a downstream 3' splice site. Thus, PTB can function as an anti-repressor molecule to counteract the splicing inhibitory activity of SRp30c.

Authors+Show Affiliations

RNA/RNP Group, Département de Microbiologie et d'Infectiologie, Faculté de Médecine et des Sciences de la Santé, Université de Sherbrooke, Sherbrooke, Québec, Canada.No 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

17548433

Citation

Paradis, Caroline, et al. "HnRNP I/PTB Can Antagonize the Splicing Repressor Activity of SRp30c." RNA (New York, N.Y.), vol. 13, no. 8, 2007, pp. 1287-300.
Paradis C, Cloutier P, Shkreta L, et al. HnRNP I/PTB can antagonize the splicing repressor activity of SRp30c. RNA. 2007;13(8):1287-300.
Paradis, C., Cloutier, P., Shkreta, L., Toutant, J., Klarskov, K., & Chabot, B. (2007). HnRNP I/PTB can antagonize the splicing repressor activity of SRp30c. RNA (New York, N.Y.), 13(8), 1287-300.
Paradis C, et al. HnRNP I/PTB Can Antagonize the Splicing Repressor Activity of SRp30c. RNA. 2007;13(8):1287-300. PubMed PMID: 17548433.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - hnRNP I/PTB can antagonize the splicing repressor activity of SRp30c. AU - Paradis,Caroline, AU - Cloutier,Philippe, AU - Shkreta,Lulzim, AU - Toutant,Johanne, AU - Klarskov,Klaus, AU - Chabot,Benoit, Y1 - 2007/06/04/ PY - 2007/6/6/pubmed PY - 2007/8/31/medline PY - 2007/6/6/entrez SP - 1287 EP - 300 JF - RNA (New York, N.Y.) JO - RNA VL - 13 IS - 8 N2 - The control of alternative pre-mRNA splicing often requires the participation of factors displaying synergistic or antagonistic activities. In the hnRNP A1 pre-mRNA, three elements promote the exclusion of alternative exon 7B, while a fourth intron element (CE9) represses splicing of exon 7B to the downstream exon. We have shown previously that the 5' portion of the 38-nucleotide-long CE9 element is bound by SRp30c, and that this interaction is important for repression in vitro. To determine whether SRp30c alone can impose repression, we tested a high-affinity SRp30c binding site that we identified using the SELEX protocol. We find that multiple high-affinity SRp30c sites are required to replicate the level of repression obtained with CE9, and that both the 5' and the 3' portions of CE9 contribute to SRp30c binding. Performing RNA affinity chromatography with the complete CE9 element recovered hnRNP I/PTB. Surprisingly however, His-tagged PTB reduced the binding of SRp30c to CE9 in a nuclear extract, stimulated splicing to a downstream 3' splice site, and relieved the CE9-mediated splicing repression in vitro. Our in vivo results are consistent with the notion that increasing PTB levels alleviates the repression imposed by CE9 to a downstream 3' splice site. Thus, PTB can function as an anti-repressor molecule to counteract the splicing inhibitory activity of SRp30c. SN - 1355-8382 UR - https://www.unboundmedicine.com/medline/citation/17548433/hnRNP_I/PTB_can_antagonize_the_splicing_repressor_activity_of_SRp30c_ L2 - http://www.rnajournal.org/cgi/pmidlookup?view=long&pmid=17548433 DB - PRIME DP - Unbound Medicine ER -