Proteomics uncovers novel components of an interactive protein network supporting RNA export in trypanosomes

Alexandre Haruo Inoue, Patricia Ferreira Domingues, Mariana Serpeloni, Priscila Mazzocchi Hiraiwa, Newton Medeiros Vidal, Erin R. Butterfield, Ricardo Canavate Del Pino, Adriana Ludwig, Cordula Boehm, Mark C. Field, Andréa Rodrigues Ávila (Lead / Corresponding author)

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)
26 Downloads (Pure)

Abstract

In trypanosomatids, transcription is polycistronic and all mRNAs are processed by trans-splicing with export mediated by non-canonical mechanisms. Although mRNA export is central to gene regulation and expression, few orthologs of proteins involved in mRNA export in higher eukaryotes are detectable in trypanosome genomes, necessitating direct identification of protein components. We previously described conserved mRNA export pathway components in Trypanosoma cruzi, including orthologs of Sub2, a component of the TREX complex, and eIF4AIII (previously Hel45), a core component of the exon junction complex (EJC). Here, we searched for protein interactors of both proteins using cryomilling and mass spectrometry. Significant overlap between TcSub2 and TceIF4AIII-interacting protein cohorts suggests that both proteins associate with similar machinery. We identified several interactions with conserved core components of the EJC and multiple additional complexes, together with proteins specific to trypanosomatids. Additional immunoisolations of kinetoplastid-specific proteins both validated and extended the super-interactome, which is capable of supporting RNA processing from splicing through to nuclear export and cytoplasmic events. We also suggest that only proteomics is powerful enough to uncover the high connectivity between multiple aspects of mRNA metabolism and to uncover kinetoplastid-specific components that create a unique amalgam to support trypanosome mRNA maturation.

Original languageEnglish
Article number100208
Number of pages25
JournalMolecular & Cellular Proteomics
Volume21
Issue number3
Early online date25 Jan 2022
DOIs
Publication statusPublished - 1 Mar 2022

Keywords

  • mRNA export
  • trypanosomes
  • gene expression
  • proteomics
  • evolution

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