Characterisation of the biflavonoid hinokiflavone as a pre-mRNA splicing modulator that inhibits SENP
- University of Dundee, United Kingdom
- WestCHEM School of Chemistry, University of Glasgow, United Kingdom
Abstract
We have identified the plant biflavonoid hinokiflavone as an inhibitor of splicing in vitro and modulator of alternative splicing in cells. Chemical synthesis confirms hinokiflavone is the active molecule. Hinokiflavone inhibits splicing in vitro by blocking spliceosome assembly, leading to accumulation of the A complex. Cells treated with hinokiflavone show altered subnuclear organization specifically of splicing factors required for A complex formation, which relocalize together with SUMO1 and SUMO2 into enlarged nuclear speckles. Hinokiflavone increases protein SUMOylation levels, both in in vitro splicing reactions and in cells. Hinokiflavone also inhibited a purified, E. coli expressed SUMO protease, SENP1, in vitro, indicating the increase in SUMOylated proteins results primarily from inhibition of de-SUMOylation. Using a quantitative proteomics assay we identified many SUMO2 sites whose levels increased in cells following hinokiflavone treatment, with the major targets including 6 proteins that are components of the U2 snRNP and required for A complex formation.
Data availability
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Characterisation of the biflavonoid hinokiflavone as a pre-mRNA splicing modulator that inhibits SENPPublicly available at PRIDE (accession no. PXD007629).
Article and author information
Author details
Funding
Wellcome (073980/Z/03/B)
- Angus I Lamond
Wellcome (097045/B/11/Z)
- Angus I Lamond
Wellcome (098391/Z/12/7)
- Ronald T Hay
Wellcome (105606/Z/14/Z)
- Ronald T Hay
European Commission (PITN-GA-2011-290257)
- Triin Tammsalu
Engineering and Physical Sciences Research Council (EP/L50497X/1)
- Lewis J King
Engineering and Physical Sciences Research Council (EP/L50497X/1)
- Helmi Kreinin
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Juan Valcárcel, Centre de Regulació Genòmica (CRG), Barcelona, Spain
Version history
- Received: April 2, 2017
- Accepted: September 6, 2017
- Accepted Manuscript published: September 8, 2017 (version 1)
- Version of Record published: September 28, 2017 (version 2)
Copyright
© 2017, Pawellek et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Characterisation of the biflavonoid hinokiflavone as a pre-mRNA splicing modulator that inhibits SENP
eLife 6:e27402.
https://doi.org/10.7554/eLife.27402
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