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Molecular design of a splicing switch responsive to the RNA binding protein Tra2β

Grellscheid, SN; Dalgliesh, C; Rozanska, A; Grellscheid, D; Bourgeois, CF; Stévenin, J; Elliott, DJ.

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Authors

C Dalgliesh

A Rozanska

D Grellscheid

CF Bourgeois

J Stévenin

DJ. Elliott



Abstract

Tra2β regulates a number of splicing switches including activation of the human testis-specific exon HIPK3-T in the Homeodomain Interacting Protein Kinase 3 gene. By testing HIPK3-T exons of different intrinsic strengths, we found Tra2β most efficiently activated splicing inclusion of intrinsically weak exons, although these were spliced at a lower overall level. Both the RRM and N-terminal RS-rich region of Tra2β were required for splicing activation. Bioinformatic searches for splicing enhancers and repressors mapped four physically distinct exonic splicing enhancers (ESEs) within HIPK3-T, each containing the known Tra2β AGAA-rich binding site. Surprisingly disruption of each single ESE prevented Tra2β-mediated activation, although single mutated exons could still bind Tra2β protein by gel shifts and functional splicing analyses. Titration experiments indicate an additive model of HIPK3-T splicing activation, requiring availability of an array of four distinct ESEs to enable splicing activation. To enable this efficient Tra2β-mediated splicing switch to operate, a closely adjacent downstream and potentially competitive stronger 5′-splice site is actively repressed. Our data indicate that a novel arrangement of multiple mono-specific AGAA-rich ESEs coupled to a weak 5′-splice site functions as a responsive gauge. This gauge monitors changes in the specific nuclear concentration of the RNA binding protein Tra2β, and co-ordinately regulates HIPK3-T exon splicing inclusion.

Citation

Grellscheid, S., Dalgliesh, C., Rozanska, A., Grellscheid, D., Bourgeois, C., Stévenin, J., & Elliott, D. (2011). Molecular design of a splicing switch responsive to the RNA binding protein Tra2β. Nucleic Acids Research, 39(18), 8092-8104. https://doi.org/10.1093/nar/gkr495

Journal Article Type Article
Publication Date Oct 1, 2011
Deposit Date Feb 11, 2013
Publicly Available Date Mar 28, 2024
Journal Nucleic Acids Research
Print ISSN 0305-1048
Electronic ISSN 1362-4962
Publisher Oxford University Press
Peer Reviewed Peer Reviewed
Volume 39
Issue 18
Pages 8092-8104
DOI https://doi.org/10.1093/nar/gkr495

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Copyright Statement
© The Author(s) 2011. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.





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