A targeted oligonucleotide enhancer of SMN2 exon 7 splicing forms competing quadruplex and protein complexes in functional conditions
A targeted oligonucleotide enhancer of SMN2 exon 7 splicing forms competing quadruplex and protein complexes in functional conditions
The use of oligonucleotides to activate the splicing of selected exons is limited by a poor understanding of the mechanisms affected. A targeted bifunctional oligonucleotide enhancer of splicing (TOES) anneals to SMN2 exon 7 and carries an exonic splicing enhancer (ESE) sequence. We show that it stimulates splicing specifically of intron 6 in the presence of repressing sequences in intron 7. Complementarity to the 5' end of exon 7 increases U2AF65 binding, but the ESE sequence is required for efficient recruitment of U2 snRNP. The ESE forms at least three coexisting discrete states: a quadruplex, a complex containing only hnRNP F/H, and a complex enriched in the activator SRSF1. Neither hnRNP H nor quadruplex formation contributes to ESE activity. The results suggest that splicing limited by weak signals can be rescued by rapid exchange of TOES oligonucleotides in various complexes and raise the possibility that SR proteins associate transiently with ESEs.
Base Sequence, Exons, Humans, Introns, Nuclear Proteins, Oligonucleotides, RNA Splice Sites, RNA Splicing, Ribonucleoprotein, U2 Small Nuclear, Ribonucleoproteins, Spliceosomes, Splicing Factor U2AF, Survival of Motor Neuron 2 Protein, Journal Article, Research Support, Non-U.S. Gov't
193-205
Smith, Lindsay D.
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Dickinson, Rachel L
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Lucas, Christian M
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Cousins, Alex
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Malygin, Alexey A
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Weldon, Carika
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Perrett, Andrew J
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Bottrill, Andrew R
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Searle, Mark S
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Burley, Glenn A
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Eperon, Ian C
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9 October 2014
Smith, Lindsay D.
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Dickinson, Rachel L
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Lucas, Christian M
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Cousins, Alex
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Malygin, Alexey A
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Weldon, Carika
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Perrett, Andrew J
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Bottrill, Andrew R
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Searle, Mark S
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Burley, Glenn A
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Eperon, Ian C
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Smith, Lindsay D., Dickinson, Rachel L, Lucas, Christian M, Cousins, Alex, Malygin, Alexey A, Weldon, Carika, Perrett, Andrew J, Bottrill, Andrew R, Searle, Mark S, Burley, Glenn A and Eperon, Ian C
(2014)
A targeted oligonucleotide enhancer of SMN2 exon 7 splicing forms competing quadruplex and protein complexes in functional conditions.
Cell Reports, 9 (1), .
(doi:10.1016/j.celrep.2014.08.051).
Abstract
The use of oligonucleotides to activate the splicing of selected exons is limited by a poor understanding of the mechanisms affected. A targeted bifunctional oligonucleotide enhancer of splicing (TOES) anneals to SMN2 exon 7 and carries an exonic splicing enhancer (ESE) sequence. We show that it stimulates splicing specifically of intron 6 in the presence of repressing sequences in intron 7. Complementarity to the 5' end of exon 7 increases U2AF65 binding, but the ESE sequence is required for efficient recruitment of U2 snRNP. The ESE forms at least three coexisting discrete states: a quadruplex, a complex containing only hnRNP F/H, and a complex enriched in the activator SRSF1. Neither hnRNP H nor quadruplex formation contributes to ESE activity. The results suggest that splicing limited by weak signals can be rescued by rapid exchange of TOES oligonucleotides in various complexes and raise the possibility that SR proteins associate transiently with ESEs.
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Accepted/In Press date: 21 August 2014
e-pub ahead of print date: 25 September 2014
Published date: 9 October 2014
Keywords:
Base Sequence, Exons, Humans, Introns, Nuclear Proteins, Oligonucleotides, RNA Splice Sites, RNA Splicing, Ribonucleoprotein, U2 Small Nuclear, Ribonucleoproteins, Spliceosomes, Splicing Factor U2AF, Survival of Motor Neuron 2 Protein, Journal Article, Research Support, Non-U.S. Gov't
Identifiers
Local EPrints ID: 420579
URI: http://eprints.soton.ac.uk/id/eprint/420579
ISSN: 2211-1247
PURE UUID: c67224d8-b23d-4c8e-ae6b-dbd82eaa3049
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Date deposited: 10 May 2018 16:30
Last modified: 15 Mar 2024 19:32
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Contributors
Author:
Lindsay D. Smith
Author:
Rachel L Dickinson
Author:
Christian M Lucas
Author:
Alex Cousins
Author:
Alexey A Malygin
Author:
Carika Weldon
Author:
Andrew J Perrett
Author:
Andrew R Bottrill
Author:
Mark S Searle
Author:
Glenn A Burley
Author:
Ian C Eperon
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