iCLIP predicts the dual splicing effects of TIA-RNA interactions.

The regulation of alternative splicing involves interactions between RNA-binding proteins and pre-mRNA positions close to the splice sites. T-cell intracellular antigen 1 (TIA1) and TIA1-like 1 (TIAL1) locally enhance exon inclusion by recruiting U1 snRNP to 5' splice sites. However, effects of...

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Main Authors: Zhen Wang, Melis Kayikci, Michael Briese, Kathi Zarnack, Nicholas M Luscombe, Gregor Rot, Blaž Zupan, Tomaž Curk, Jernej Ule
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2010-10-01
Series:PLoS Biology
Online Access:https://journals.plos.org/plosbiology/article/file?id=10.1371/journal.pbio.1000530&type=printable
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author Zhen Wang
Melis Kayikci
Michael Briese
Kathi Zarnack
Nicholas M Luscombe
Gregor Rot
Blaž Zupan
Tomaž Curk
Jernej Ule
author_facet Zhen Wang
Melis Kayikci
Michael Briese
Kathi Zarnack
Nicholas M Luscombe
Gregor Rot
Blaž Zupan
Tomaž Curk
Jernej Ule
author_sort Zhen Wang
collection DOAJ
description The regulation of alternative splicing involves interactions between RNA-binding proteins and pre-mRNA positions close to the splice sites. T-cell intracellular antigen 1 (TIA1) and TIA1-like 1 (TIAL1) locally enhance exon inclusion by recruiting U1 snRNP to 5' splice sites. However, effects of TIA proteins on splicing of distal exons have not yet been explored. We used UV-crosslinking and immunoprecipitation (iCLIP) to find that TIA1 and TIAL1 bind at the same positions on human RNAs. Binding downstream of 5' splice sites was used to predict the effects of TIA proteins in enhancing inclusion of proximal exons and silencing inclusion of distal exons. The predictions were validated in an unbiased manner using splice-junction microarrays, RT-PCR, and minigene constructs, which showed that TIA proteins maintain splicing fidelity and regulate alternative splicing by binding exclusively downstream of 5' splice sites. Surprisingly, TIA binding at 5' splice sites silenced distal cassette and variable-length exons without binding in proximity to the regulated alternative 3' splice sites. Using transcriptome-wide high-resolution mapping of TIA-RNA interactions we evaluated the distal splicing effects of TIA proteins. These data are consistent with a model where TIA proteins shorten the time available for definition of an alternative exon by enhancing recognition of the preceding 5' splice site. Thus, our findings indicate that changes in splicing kinetics could mediate the distal regulation of alternative splicing.
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spelling doaj-art-a8d5a0e896eb4e3c9d1ffb8f295dfa6b2025-08-20T02:31:51ZengPublic Library of Science (PLoS)PLoS Biology1544-91731545-78852010-10-01810e100053010.1371/journal.pbio.1000530iCLIP predicts the dual splicing effects of TIA-RNA interactions.Zhen WangMelis KayikciMichael BrieseKathi ZarnackNicholas M LuscombeGregor RotBlaž ZupanTomaž CurkJernej UleThe regulation of alternative splicing involves interactions between RNA-binding proteins and pre-mRNA positions close to the splice sites. T-cell intracellular antigen 1 (TIA1) and TIA1-like 1 (TIAL1) locally enhance exon inclusion by recruiting U1 snRNP to 5' splice sites. However, effects of TIA proteins on splicing of distal exons have not yet been explored. We used UV-crosslinking and immunoprecipitation (iCLIP) to find that TIA1 and TIAL1 bind at the same positions on human RNAs. Binding downstream of 5' splice sites was used to predict the effects of TIA proteins in enhancing inclusion of proximal exons and silencing inclusion of distal exons. The predictions were validated in an unbiased manner using splice-junction microarrays, RT-PCR, and minigene constructs, which showed that TIA proteins maintain splicing fidelity and regulate alternative splicing by binding exclusively downstream of 5' splice sites. Surprisingly, TIA binding at 5' splice sites silenced distal cassette and variable-length exons without binding in proximity to the regulated alternative 3' splice sites. Using transcriptome-wide high-resolution mapping of TIA-RNA interactions we evaluated the distal splicing effects of TIA proteins. These data are consistent with a model where TIA proteins shorten the time available for definition of an alternative exon by enhancing recognition of the preceding 5' splice site. Thus, our findings indicate that changes in splicing kinetics could mediate the distal regulation of alternative splicing.https://journals.plos.org/plosbiology/article/file?id=10.1371/journal.pbio.1000530&type=printable
spellingShingle Zhen Wang
Melis Kayikci
Michael Briese
Kathi Zarnack
Nicholas M Luscombe
Gregor Rot
Blaž Zupan
Tomaž Curk
Jernej Ule
iCLIP predicts the dual splicing effects of TIA-RNA interactions.
PLoS Biology
title iCLIP predicts the dual splicing effects of TIA-RNA interactions.
title_full iCLIP predicts the dual splicing effects of TIA-RNA interactions.
title_fullStr iCLIP predicts the dual splicing effects of TIA-RNA interactions.
title_full_unstemmed iCLIP predicts the dual splicing effects of TIA-RNA interactions.
title_short iCLIP predicts the dual splicing effects of TIA-RNA interactions.
title_sort iclip predicts the dual splicing effects of tia rna interactions
url https://journals.plos.org/plosbiology/article/file?id=10.1371/journal.pbio.1000530&type=printable
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