Cross-linking mass spectrometric analysis of the endogenous TREX complex from Saccharomyces cerevisiae
- 1Institute of Biochemistry, FB08, Justus Liebig University, 35392 Giessen, Germany
- 2Institute of Biochemistry and Biology, Department of Biochemistry, University of Potsdam, 14476 Potsdam-Golm, Germany
- 3Department of Biology, Institute of Molecular Systems Biology, ETH Zurich, 8093 Zurich, Switzerland
- 4Cardio-Pulmonary Institute (CPI), EXC 2026, 35392 Giessen, Germany
- Corresponding authors: wolfgang.wende{at}chemie.bio.uni-giessen.de, leitner{at}imsb.biol.ethz.ch, katja.straesser{at}chemie.bio.uni-giessen.de
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↵5 These authors contributed equally to this work.
Abstract
The conserved TREX complex has multiple functions in gene expression such as transcription elongation, 3′ end processing, mRNP assembly and nuclear mRNA export as well as the maintenance of genomic stability. In Saccharomyces cerevisiae, TREX is composed of the pentameric THO complex, the DEAD-box RNA helicase Sub2, the nuclear mRNA export adaptor Yra1, and the SR-like proteins Gbp2 and Hrb1. Here, we present the structural analysis of the endogenous TREX complex of S. cerevisiae purified from its native environment. To this end, we used cross-linking mass spectrometry to gain structural information on regions of the complex that are not accessible to classical structural biology techniques. We also used negative-stain electron microscopy to investigate the organization of the cross-linked complex used for XL-MS by comparing our endogenous TREX complex with recently published structural models of recombinant THO–Sub2 complexes. According to our analysis, the endogenous yeast TREX complex preferentially assembles into a dimer.
Keywords
- Received June 23, 2023.
- Accepted August 22, 2023.
This article, published in RNA, is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.
