We identify proteins that associate with the THO core complex, and show that the TEX1 and MOS11 components functionally interact, affecting mRNA export and splicing as well as plant development.
TREX (TRanscription-EXport) is a multiprotein complex that plays a central role in the coordination of synthesis, processing and nuclear export of mRNAs. Using targeted proteomics, we identified proteins that associate with the THO core complex of Arabidopsis TREX. In addition to the RNA helicase UAP56 and the mRNA export factors ALY2-4 and MOS11 we detected interactions with the mRNA export complex TREX-2 and multiple spliceosomal components. Plants defective in the THO component TEX1 or in the mRNA export factor MOS11 (orthologue of human CIP29) are mildly affected. However, tex1 mos11 double-mutant plants show marked defects in vegetative and reproductive development. In tex1 plants, the levels of tasiRNAs are reduced, while miR173 levels are decreased in mos11 mutants. In nuclei of mos11 cells increased mRNA accumulation was observed, while no mRNA export defect was detected with tex1 cells. Nevertheless, in tex1 mos11 double-mutants, the mRNA export defect was clearly enhanced relative to mos11. The subnuclear distribution of TEX1 substantially overlaps with that of splicing-related SR proteins and in tex1 plants the ratio of certain alternative splicing events is altered. Our results demonstrate that Arabidopsis TEX1 and MOS11 are involved in distinct steps of the biogenesis of mRNAs and small RNAs, and that they interact regarding some aspects, but act independently in others.
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We thank Jelle Van Leene and Geert De Jaeger for advice regarding cell culture transformation and for providing Arabidopsis PSB-D cells, Liang-zi Zhou for help with the protoplast transformation experiment, Eduard Hochmuth for recording mass spectra, Patrick Motte for plasmids pBI35S::atRSZp22-mRFP1 and pBI35S::atRSZp33-mRFP1, Claus Schwechheimer for pid-14 seeds, the Nottingham Arabidopsis Stock Centre (NASC) for providing Arabidopsis T-DNA insertion lines. This work was supported by the German Research Foundation (DFG) through grant SFB960 to M. G and K. D. G.
BBS, MG, KDG conceived and designed the experiments. BBS, HFE, SE, AP, TS, MM, MG performed the experiments. BBS, HFE, SE, AP, AB, GM, RM, TS, GL, MM, MG, KDG analysed the data. JH, GM, RM, TS, GL, MM, MG, KDG contributed reagents/materials/analysis tools. MG, KDG wrote the paper. All authors reviewed the results and approved the final version of the manuscript.
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Sørensen, B.B., Ehrnsberger, H.F., Esposito, S. et al. The Arabidopsis THO/TREX component TEX1 functionally interacts with MOS11 and modulates mRNA export and alternative splicing events. Plant Mol Biol 93, 283–298 (2017). https://doi.org/10.1007/s11103-016-0561-9
- mRNA processing/export
- Protein complexes
- RNA polymerase II