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

Andrea Pawellek, Ursula Ryder, Triin Tammsalu, Lewis J King, Helmi Kreinin, Tony Ly, Ronald T Hay, Richard C Hartley, Angus I Lamond

Research output: Contribution to journalArticlepeer-review

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, preventing formation of the B 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 containing polyadenylated RNA. Hinokiflavone increases protein SUMOylation levels, both in in vitro splicing reactions and in cells. Hinokiflavone also inhibited a purified,E. coliexpressed 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 six proteins that are components of the U2 snRNP and required for A complex formation.

Original languageEnglish
Article numbere27402
Number of pages36
JournaleLIFE
Volume6
DOIs
Publication statusPublished - 8 Sep 2017

Keywords

  • RNA splicing
  • SENP1
  • SUMO
  • U2 snRNP
  • biochemistry
  • hinokiflavone
  • none

Fingerprint

Dive into the research topics of 'Characterisation of the biflavonoid hinokiflavone as a pre-mRNA splicing modulator that inhibits SENP'. Together they form a unique fingerprint.

Cite this