hnRNP A1 and hnRNP F modulate the alternative splicing of exon 11 of the insulin receptor gene.
Exon 11 of the insulin receptor gene (INSR) is alternatively spliced in a developmentally and tissue-specific manner. Linker scanning mutations in a 5' GA-rich enhancer in intron 10 identified AGGGA sequences that are important for enhancer function. Using RNA-affinity purification and mass spe...
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doaj-76126059c4984455833b50e99812271f2020-11-25T01:56:04ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-01611e2786910.1371/journal.pone.0027869hnRNP A1 and hnRNP F modulate the alternative splicing of exon 11 of the insulin receptor gene.Indrani TalukdarSupriya SenRodolfo UrbanoJames ThompsonJohn R YatesNicholas J G WebsterExon 11 of the insulin receptor gene (INSR) is alternatively spliced in a developmentally and tissue-specific manner. Linker scanning mutations in a 5' GA-rich enhancer in intron 10 identified AGGGA sequences that are important for enhancer function. Using RNA-affinity purification and mass spectrometry, we identified hnRNP F and hnRNP A1 binding to these AGGGA sites and also to similar motifs at the 3' end of the intron. The hnRNPs have opposite functional effects with hnRNP F promoting and hnRNP A1 inhibiting exon 11 inclusion, and deletion of the GA-rich elements eliminates both effects. We also observed specific binding of hnRNP A1 to the 5' splice site of intron 11. The SR protein SRSF1 (SF2/ASF) co-purified on the GA-rich enhancer and, interestingly, also competes with hnRNP A1 for binding to the splice site. A point mutation -3U→C decreases hnRNP A1 binding, increases SRSF1 binding and renders the exon constitutive. Lastly, our data point to a functional interaction between hnRNP F and SRSF1 as a mutant that eliminates SRSF1 binding to exon 11, or a SRSF1 knockdown, which prevents the stimulatory effect of hnRNP F over expression.http://europepmc.org/articles/PMC3223206?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Indrani Talukdar Supriya Sen Rodolfo Urbano James Thompson John R Yates Nicholas J G Webster |
spellingShingle |
Indrani Talukdar Supriya Sen Rodolfo Urbano James Thompson John R Yates Nicholas J G Webster hnRNP A1 and hnRNP F modulate the alternative splicing of exon 11 of the insulin receptor gene. PLoS ONE |
author_facet |
Indrani Talukdar Supriya Sen Rodolfo Urbano James Thompson John R Yates Nicholas J G Webster |
author_sort |
Indrani Talukdar |
title |
hnRNP A1 and hnRNP F modulate the alternative splicing of exon 11 of the insulin receptor gene. |
title_short |
hnRNP A1 and hnRNP F modulate the alternative splicing of exon 11 of the insulin receptor gene. |
title_full |
hnRNP A1 and hnRNP F modulate the alternative splicing of exon 11 of the insulin receptor gene. |
title_fullStr |
hnRNP A1 and hnRNP F modulate the alternative splicing of exon 11 of the insulin receptor gene. |
title_full_unstemmed |
hnRNP A1 and hnRNP F modulate the alternative splicing of exon 11 of the insulin receptor gene. |
title_sort |
hnrnp a1 and hnrnp f modulate the alternative splicing of exon 11 of the insulin receptor gene. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2011-01-01 |
description |
Exon 11 of the insulin receptor gene (INSR) is alternatively spliced in a developmentally and tissue-specific manner. Linker scanning mutations in a 5' GA-rich enhancer in intron 10 identified AGGGA sequences that are important for enhancer function. Using RNA-affinity purification and mass spectrometry, we identified hnRNP F and hnRNP A1 binding to these AGGGA sites and also to similar motifs at the 3' end of the intron. The hnRNPs have opposite functional effects with hnRNP F promoting and hnRNP A1 inhibiting exon 11 inclusion, and deletion of the GA-rich elements eliminates both effects. We also observed specific binding of hnRNP A1 to the 5' splice site of intron 11. The SR protein SRSF1 (SF2/ASF) co-purified on the GA-rich enhancer and, interestingly, also competes with hnRNP A1 for binding to the splice site. A point mutation -3U→C decreases hnRNP A1 binding, increases SRSF1 binding and renders the exon constitutive. Lastly, our data point to a functional interaction between hnRNP F and SRSF1 as a mutant that eliminates SRSF1 binding to exon 11, or a SRSF1 knockdown, which prevents the stimulatory effect of hnRNP F over expression. |
url |
http://europepmc.org/articles/PMC3223206?pdf=render |
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