Characterisation of the biflavonoid hinokiflavone as a pre-mRNA splicing modulator that inhibits SENP

Pawellek, A., Ryder, U., Tammsalu, T., King, L. J., Kreinin, H., Ly, T., Hay, R. T., Hartley, R. and Lamond, A. I. (2017) Characterisation of the biflavonoid hinokiflavone as a pre-mRNA splicing modulator that inhibits SENP. eLife, 6, e27402. (doi:10.7554/eLife.27402) (PMID:28884683)

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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.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:King, Mr Lewis and Kreinin, Miss Helmi and Hartley, Professor Richard
Authors: Pawellek, A., Ryder, U., Tammsalu, T., King, L. J., Kreinin, H., Ly, T., Hay, R. T., Hartley, R., and Lamond, A. I.
College/School:College of Science and Engineering > School of Chemistry
Journal Name:eLife
Publisher:eLife Sciences Publications
ISSN:2050-084X
ISSN (Online):2050-084X
Copyright Holders:Copyright © 2017 Pawellek et al.
First Published:First published in eLife 6:e27402
Publisher Policy:Reproduced under a Creative Commons License

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
642131Doctoral Training Grant 2013 - 2017Mary Beth KneafseyEngineering and Physical Sciences Research Council (EPSRC)EP/L50497X/1RSI - RESEARCH STRATEGY & INNOVATION