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 |
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Status: | Published |
Refereed: | Yes |
Glasgow Author(s) Enlighten ID: | Hartley, Professor Richard and King, Mr Lewis and Kreinin, Miss Helmi |
Creator Roles: | Hartley, R.Funding acquisition, Supervision, Writing – review and editing King, L.Investigation, Methodology, Resources Kreinin, H.Investigation, Methodology, Resources |
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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