The Experts below are selected from a list of 273 Experts worldwide ranked by ideXlab platform
Alberto R Kornblihtt - One of the best experts on this subject based on the ideXlab platform.
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a slow rna polymerase ii affects alternative splicing in vivo
Molecular Cell, 2003Co-Authors: Manuel De La Mata, Paula Cramer, Sebastian Kadener, Claudio R Alonso, Juan Pablo Fededa, Matias Blaustein, Federico Pelisch, David Bentley, Alberto R KornblihttAbstract:Changes in Promoter Structure and occupation have been shown to modify the splicing pattern of several genes, evidencing a coupling between transcription and alternative splicing. It has been proposed that the Promoter effect involves modulation of RNA pol II elongation rates. The C4 point mutation of the Drosophila pol II largest subunit confers on the enzyme a lower elongation rate. Here we show that expression of a human equivalent to Drosophila's C4 pol II in human cultured cells affects alternative splicing of the fibronectin EDI exon and adenovirus E1a pre-mRNA. Most importantly, resplicing of the Hox gene Ultrabithorax is stimulated in Drosophila embryos mutant for C4, which demonstrates the transcriptional control of alternative splicing on an endogenous gene. These results provide a direct proof for the elongation control of alternative splicing in vivo.
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regulation of alternative splicing by a transcriptional enhancer through rna pol ii elongation
Proceedings of the National Academy of Sciences of the United States of America, 2002Co-Authors: Sebastian Kadener, Juan Pablo Fededa, Michael Rosbash, Alberto R KornblihttAbstract:Promoters and enhancers are cis-acting elements that control gene transcription via complex networks of protein–DNA and protein–protein 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.
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coupling of transcription with alternative splicing rna pol ii Promoters modulate sf2 asf and 9g8 effects on an exonic splicing enhancer
Molecular Cell, 1999Co-Authors: Paula Cramer, Javier F Caceres, Demian Cazalla, Sebastian Kadener, Andres F Muro, Francisco E Baralle, Alberto R KornblihttAbstract:Abstract Alternative mRNA splicing of the fibronectin EDI exon is controlled by a purine-rich exonic splicing enhancer (ESE), postulated as a binding site for SR proteins. By using a transient expression alternative splicing assay combined with Promoter swapping, we have demonstrated that the Promoter can also control EDI splicing, arguing for coupling between the transcription and splicing machineries. We now report that the SR proteins SF2/ASF and 9G8 stimulate EDI splicing in vivo and that their effect requires an intact EDI ESE. Most importantly, we show that sensitivity to these SR proteins critically depends on the Promoter Structure, suggesting that the transcription machinery modulates their recruitment to the ESE.
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functional association between Promoter Structure and transcript alternative splicing
Proceedings of the National Academy of Sciences of the United States of America, 1997Co-Authors: Paula Cramer, C G Pesce, F E Baralle, Alberto R KornblihttAbstract:It has been assumed that constitutive and regulated splicing of RNA polymerase II transcripts depends exclusively on signals present in the RNA molecule. Here we show that changes in Promoter Structure strongly affect splice site selection. We investigated the splicing of the ED I exon, which encodes a facultative type III repeat of fibronectin, whose inclusion is regulated during development and in proliferative processes. We used an alternative splicing assay combined with Promoter swapping to demonstrate that the extent of ED I splicing is dependent on the Promoter Structure from which the transcript originated and that this regulation is independent of the Promoter strength. Thus, these results provide the first evidence for coupling between alternative splicing and Promoter-specific transcription, which agrees with recent cytological and biochemical evidence of coordination between splicing and transcription.
Sebastian Kadener - One of the best experts on this subject based on the ideXlab platform.
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a slow rna polymerase ii affects alternative splicing in vivo
Molecular Cell, 2003Co-Authors: Manuel De La Mata, Paula Cramer, Sebastian Kadener, Claudio R Alonso, Juan Pablo Fededa, Matias Blaustein, Federico Pelisch, David Bentley, Alberto R KornblihttAbstract:Changes in Promoter Structure and occupation have been shown to modify the splicing pattern of several genes, evidencing a coupling between transcription and alternative splicing. It has been proposed that the Promoter effect involves modulation of RNA pol II elongation rates. The C4 point mutation of the Drosophila pol II largest subunit confers on the enzyme a lower elongation rate. Here we show that expression of a human equivalent to Drosophila's C4 pol II in human cultured cells affects alternative splicing of the fibronectin EDI exon and adenovirus E1a pre-mRNA. Most importantly, resplicing of the Hox gene Ultrabithorax is stimulated in Drosophila embryos mutant for C4, which demonstrates the transcriptional control of alternative splicing on an endogenous gene. These results provide a direct proof for the elongation control of alternative splicing in vivo.
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regulation of alternative splicing by a transcriptional enhancer through rna pol ii elongation
Proceedings of the National Academy of Sciences of the United States of America, 2002Co-Authors: Sebastian Kadener, Juan Pablo Fededa, Michael Rosbash, Alberto R KornblihttAbstract:Promoters and enhancers are cis-acting elements that control gene transcription via complex networks of protein–DNA and protein–protein 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.
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coupling of transcription with alternative splicing rna pol ii Promoters modulate sf2 asf and 9g8 effects on an exonic splicing enhancer
Molecular Cell, 1999Co-Authors: Paula Cramer, Javier F Caceres, Demian Cazalla, Sebastian Kadener, Andres F Muro, Francisco E Baralle, Alberto R KornblihttAbstract:Abstract Alternative mRNA splicing of the fibronectin EDI exon is controlled by a purine-rich exonic splicing enhancer (ESE), postulated as a binding site for SR proteins. By using a transient expression alternative splicing assay combined with Promoter swapping, we have demonstrated that the Promoter can also control EDI splicing, arguing for coupling between the transcription and splicing machineries. We now report that the SR proteins SF2/ASF and 9G8 stimulate EDI splicing in vivo and that their effect requires an intact EDI ESE. Most importantly, we show that sensitivity to these SR proteins critically depends on the Promoter Structure, suggesting that the transcription machinery modulates their recruitment to the ESE.
Paula Cramer - One of the best experts on this subject based on the ideXlab platform.
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a slow rna polymerase ii affects alternative splicing in vivo
Molecular Cell, 2003Co-Authors: Manuel De La Mata, Paula Cramer, Sebastian Kadener, Claudio R Alonso, Juan Pablo Fededa, Matias Blaustein, Federico Pelisch, David Bentley, Alberto R KornblihttAbstract:Changes in Promoter Structure and occupation have been shown to modify the splicing pattern of several genes, evidencing a coupling between transcription and alternative splicing. It has been proposed that the Promoter effect involves modulation of RNA pol II elongation rates. The C4 point mutation of the Drosophila pol II largest subunit confers on the enzyme a lower elongation rate. Here we show that expression of a human equivalent to Drosophila's C4 pol II in human cultured cells affects alternative splicing of the fibronectin EDI exon and adenovirus E1a pre-mRNA. Most importantly, resplicing of the Hox gene Ultrabithorax is stimulated in Drosophila embryos mutant for C4, which demonstrates the transcriptional control of alternative splicing on an endogenous gene. These results provide a direct proof for the elongation control of alternative splicing in vivo.
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coupling of transcription with alternative splicing rna pol ii Promoters modulate sf2 asf and 9g8 effects on an exonic splicing enhancer
Molecular Cell, 1999Co-Authors: Paula Cramer, Javier F Caceres, Demian Cazalla, Sebastian Kadener, Andres F Muro, Francisco E Baralle, Alberto R KornblihttAbstract:Abstract Alternative mRNA splicing of the fibronectin EDI exon is controlled by a purine-rich exonic splicing enhancer (ESE), postulated as a binding site for SR proteins. By using a transient expression alternative splicing assay combined with Promoter swapping, we have demonstrated that the Promoter can also control EDI splicing, arguing for coupling between the transcription and splicing machineries. We now report that the SR proteins SF2/ASF and 9G8 stimulate EDI splicing in vivo and that their effect requires an intact EDI ESE. Most importantly, we show that sensitivity to these SR proteins critically depends on the Promoter Structure, suggesting that the transcription machinery modulates their recruitment to the ESE.
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functional association between Promoter Structure and transcript alternative splicing
Proceedings of the National Academy of Sciences of the United States of America, 1997Co-Authors: Paula Cramer, C G Pesce, F E Baralle, Alberto R KornblihttAbstract:It has been assumed that constitutive and regulated splicing of RNA polymerase II transcripts depends exclusively on signals present in the RNA molecule. Here we show that changes in Promoter Structure strongly affect splice site selection. We investigated the splicing of the ED I exon, which encodes a facultative type III repeat of fibronectin, whose inclusion is regulated during development and in proliferative processes. We used an alternative splicing assay combined with Promoter swapping to demonstrate that the extent of ED I splicing is dependent on the Promoter Structure from which the transcript originated and that this regulation is independent of the Promoter strength. Thus, these results provide the first evidence for coupling between alternative splicing and Promoter-specific transcription, which agrees with recent cytological and biochemical evidence of coordination between splicing and transcription.
Juan Pablo Fededa - One of the best experts on this subject based on the ideXlab platform.
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a slow rna polymerase ii affects alternative splicing in vivo
Molecular Cell, 2003Co-Authors: Manuel De La Mata, Paula Cramer, Sebastian Kadener, Claudio R Alonso, Juan Pablo Fededa, Matias Blaustein, Federico Pelisch, David Bentley, Alberto R KornblihttAbstract:Changes in Promoter Structure and occupation have been shown to modify the splicing pattern of several genes, evidencing a coupling between transcription and alternative splicing. It has been proposed that the Promoter effect involves modulation of RNA pol II elongation rates. The C4 point mutation of the Drosophila pol II largest subunit confers on the enzyme a lower elongation rate. Here we show that expression of a human equivalent to Drosophila's C4 pol II in human cultured cells affects alternative splicing of the fibronectin EDI exon and adenovirus E1a pre-mRNA. Most importantly, resplicing of the Hox gene Ultrabithorax is stimulated in Drosophila embryos mutant for C4, which demonstrates the transcriptional control of alternative splicing on an endogenous gene. These results provide a direct proof for the elongation control of alternative splicing in vivo.
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regulation of alternative splicing by a transcriptional enhancer through rna pol ii elongation
Proceedings of the National Academy of Sciences of the United States of America, 2002Co-Authors: Sebastian Kadener, Juan Pablo Fededa, Michael Rosbash, Alberto R KornblihttAbstract:Promoters and enhancers are cis-acting elements that control gene transcription via complex networks of protein–DNA and protein–protein 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.
Kiyoto Motojima - One of the best experts on this subject based on the ideXlab platform.
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dual Promoter Structure of mouse and human fatty acid translocase cd36 genes and unique transcriptional activation by peroxisome proliferator activated receptor α and γ ligands
Journal of Biological Chemistry, 2002Co-Authors: Osamu Sato, Chikako Kuriki, Yuka Fukui, Kiyoto MotojimaAbstract:Fatty acid translocase (FAT)/CD36 is a glycoprotein involved in multiple membrane functions including uptake of long-chain fatty acids and oxidized low density lipoprotein. In mice, expression of the gene is regulated by peroxisome proliferator-activated receptor (PPAR) α in the liver and by PPARγ in the adipose tissues (Motojima, K., Passilly, P. P., Peters, J. M., Gonzalez, F. J., and Latruffe, N. (1998) J. Biol. Chem. 273, 16710–16714). However, the time course of PPARα ligand-induced expression of FAT/CD36 in the liver, and also in the cultured hepatoma cells, is significantly slower than those of other PPARα target genes. To study the molecular mechanism of the slow transcriptional activation of the gene by a PPAR ligand, we first cloned the 5′ ends of the mRNA and then the mouse gene Promoter region from a genomic bacterial artificial chromosome library. Sequencing analyses showed that transcription of the gene starts at two initiation sites 16 kb apart and splicing occurs alternatively, producing at least three mRNA species with different 5′-noncoding regions. The PPARα ligand-responsive Promoter in the liver was identified as the new upstream Promoter where we found several possible binding sites for lipid metabolism-related transcriptional factors but not for PPAR. Neither Promoter responded to a PPARα ligand in thein vitro or in vivo reporter assays using cultured hepatoma cells and the liver of living mice. We also have cloned the human FAT/CD36 gene from a bacterial artificial chromosome library and identified a new independent Promoter that is located 13 kb upstream of the previously reported Promoter. Only the upstream Promoter responded to PPARα and PPARγ ligands in a cell type-specific manner. The absence of PPRE in the responding upstream Promoter region, the delayed activation by the ligand, and the results of the reporter assays all suggested that transcriptional activation of the FAT/CD36 gene by PPAR ligands is indirectly dependent on PPAR.
