Regulation of alternative splicing by a transcriptional enhancer through RNA pol II elongation
Promoters and enhancers are cis-acting elements that control gene transcription via complex networks of protein-DNA and proteinprotein interactions. Whereas promoters deal with putting in place the RNA polymerase, both enhancers and promoters can control transcriptional initiation and elongation. We...
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2002
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00278424_v99_n12_p8185_Kadener http://hdl.handle.net/20.500.12110/paper_00278424_v99_n12_p8185_Kadener |
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paper:paper_00278424_v99_n12_p8185_Kadener2023-06-08T14:54:38Z Regulation of alternative splicing by a transcriptional enhancer through RNA pol II elongation Kadener, Sebastián Fededa, Juan Pablo Kornblihtt, Alberto Rodolfo Coupling RNA processing Transcription alpha globin cis acting element fibronectin RNA polymerase II alternative RNA splicing article chromatin enhancer region exon genetic transcription genetic transfection immunoprecipitation in vivo study priority journal promoter region protein DNA interaction protein protein interaction RNA processing Simian virus 40 transcription initiation transcription regulation Alternative Splicing Animals Base Sequence Cell Line DNA Primers Enhancer Elements (Genetics) Exons Fibronectins Gene Expression Regulation Mammals Models, Genetic Polymerase Chain Reaction RNA Polymerase II Simian virus 40 Transcription, Genetic Transfection Mammalia Simiae Simian virus 40 Promoters and enhancers are cis-acting elements that control gene transcription via complex networks of protein-DNA and proteinprotein interactions. Whereas promoters deal with putting in place the RNA polymerase, both enhancers and promoters can control transcriptional initiation and elongation. We have previously shown that promoter structure modulates alternative splicing, strengthening the concept of a physical and functional coupling between transcription and splicing. Here we report that the promoter effect is due to the control of RNA pol II elongation. We found that the simian virus 40 (SV40) transcriptional enhancer, inserted in fibronectin (FN) minigene constructs transfected into mammalian cells, controls alternative splicing by inhibiting inclusion of the FN extra domain I (EDI) exon into mature mRNA. Deletion analysis of enhancer subdomains and competitions in vivo with excess of specific enhancer DNA subfragments demonstrate that the "minimal" enhancer, consisting of two 72-bp repeats, is responsible for the splicing effect. The 72-bp repeat region has been reported to promote RNA pol II elongation. When transcription is driven by the α-globin promoter linked to the SV40 enhancer, basal EDI inclusion and activation by the SR (Ser-Arg-rich) protein SF2/ASF are much lower than with other promoters. Deletion of only one of the two 72-bp repeats not only provokes higher EDI inclusion levels but allows responsiveness to SF2/ASF. These effects are the consequence of a decrease in RNA pol II elongation evidenced both by an increase in the proportions of shorter proximal over full length transcripts and by higher pol II densities upstream of the alternative exon detected by chromatin immunoprecipitation. Fil:Kadener, S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Fededa, J.P. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Kornblihtt, A.R. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2002 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00278424_v99_n12_p8185_Kadener http://hdl.handle.net/20.500.12110/paper_00278424_v99_n12_p8185_Kadener |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Coupling RNA processing Transcription alpha globin cis acting element fibronectin RNA polymerase II alternative RNA splicing article chromatin enhancer region exon genetic transcription genetic transfection immunoprecipitation in vivo study priority journal promoter region protein DNA interaction protein protein interaction RNA processing Simian virus 40 transcription initiation transcription regulation Alternative Splicing Animals Base Sequence Cell Line DNA Primers Enhancer Elements (Genetics) Exons Fibronectins Gene Expression Regulation Mammals Models, Genetic Polymerase Chain Reaction RNA Polymerase II Simian virus 40 Transcription, Genetic Transfection Mammalia Simiae Simian virus 40 |
| spellingShingle |
Coupling RNA processing Transcription alpha globin cis acting element fibronectin RNA polymerase II alternative RNA splicing article chromatin enhancer region exon genetic transcription genetic transfection immunoprecipitation in vivo study priority journal promoter region protein DNA interaction protein protein interaction RNA processing Simian virus 40 transcription initiation transcription regulation Alternative Splicing Animals Base Sequence Cell Line DNA Primers Enhancer Elements (Genetics) Exons Fibronectins Gene Expression Regulation Mammals Models, Genetic Polymerase Chain Reaction RNA Polymerase II Simian virus 40 Transcription, Genetic Transfection Mammalia Simiae Simian virus 40 Kadener, Sebastián Fededa, Juan Pablo Kornblihtt, Alberto Rodolfo Regulation of alternative splicing by a transcriptional enhancer through RNA pol II elongation |
| topic_facet |
Coupling RNA processing Transcription alpha globin cis acting element fibronectin RNA polymerase II alternative RNA splicing article chromatin enhancer region exon genetic transcription genetic transfection immunoprecipitation in vivo study priority journal promoter region protein DNA interaction protein protein interaction RNA processing Simian virus 40 transcription initiation transcription regulation Alternative Splicing Animals Base Sequence Cell Line DNA Primers Enhancer Elements (Genetics) Exons Fibronectins Gene Expression Regulation Mammals Models, Genetic Polymerase Chain Reaction RNA Polymerase II Simian virus 40 Transcription, Genetic Transfection Mammalia Simiae Simian virus 40 |
| description |
Promoters and enhancers are cis-acting elements that control gene transcription via complex networks of protein-DNA and proteinprotein interactions. Whereas promoters deal with putting in place the RNA polymerase, both enhancers and promoters can control transcriptional initiation and elongation. We have previously shown that promoter structure modulates alternative splicing, strengthening the concept of a physical and functional coupling between transcription and splicing. Here we report that the promoter effect is due to the control of RNA pol II elongation. We found that the simian virus 40 (SV40) transcriptional enhancer, inserted in fibronectin (FN) minigene constructs transfected into mammalian cells, controls alternative splicing by inhibiting inclusion of the FN extra domain I (EDI) exon into mature mRNA. Deletion analysis of enhancer subdomains and competitions in vivo with excess of specific enhancer DNA subfragments demonstrate that the "minimal" enhancer, consisting of two 72-bp repeats, is responsible for the splicing effect. The 72-bp repeat region has been reported to promote RNA pol II elongation. When transcription is driven by the α-globin promoter linked to the SV40 enhancer, basal EDI inclusion and activation by the SR (Ser-Arg-rich) protein SF2/ASF are much lower than with other promoters. Deletion of only one of the two 72-bp repeats not only provokes higher EDI inclusion levels but allows responsiveness to SF2/ASF. These effects are the consequence of a decrease in RNA pol II elongation evidenced both by an increase in the proportions of shorter proximal over full length transcripts and by higher pol II densities upstream of the alternative exon detected by chromatin immunoprecipitation. |
| author |
Kadener, Sebastián Fededa, Juan Pablo Kornblihtt, Alberto Rodolfo |
| author_facet |
Kadener, Sebastián Fededa, Juan Pablo Kornblihtt, Alberto Rodolfo |
| author_sort |
Kadener, Sebastián |
| title |
Regulation of alternative splicing by a transcriptional enhancer through RNA pol II elongation |
| title_short |
Regulation of alternative splicing by a transcriptional enhancer through RNA pol II elongation |
| title_full |
Regulation of alternative splicing by a transcriptional enhancer through RNA pol II elongation |
| title_fullStr |
Regulation of alternative splicing by a transcriptional enhancer through RNA pol II elongation |
| title_full_unstemmed |
Regulation of alternative splicing by a transcriptional enhancer through RNA pol II elongation |
| title_sort |
regulation of alternative splicing by a transcriptional enhancer through rna pol ii elongation |
| publishDate |
2002 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00278424_v99_n12_p8185_Kadener http://hdl.handle.net/20.500.12110/paper_00278424_v99_n12_p8185_Kadener |
| work_keys_str_mv |
AT kadenersebastian regulationofalternativesplicingbyatranscriptionalenhancerthroughrnapoliielongation AT fededajuanpablo regulationofalternativesplicingbyatranscriptionalenhancerthroughrnapoliielongation AT kornblihttalbertorodolfo regulationofalternativesplicingbyatranscriptionalenhancerthroughrnapoliielongation |
| _version_ |
1768546148770906112 |