Proteomics reveal cap-dependent translation inhibitors remodel the translation machinery and translatome

J. J.David Ho, Tyler A. Cunningham, Paola Manara, Caroline A. Coughlin, Artavazd Arumov, Evan R. Roberts, Ashanti Osteen, Preet Kumar, Daniel Bilbao, Jonathan R. Krieger, Stephen Lee, Jonathan H. Schatz

Research output: Contribution to journalArticlepeer-review

Abstract

Tactical disruption of protein synthesis is an attractive therapeutic strategy, with the first-in-class eIF4A-targeting compound zotatifin in clinical evaluation for cancer and COVID-19. The full cellular impact and mechanisms of these potent molecules are undefined at a proteomic level. Here, we report mass spectrometry analysis of translational reprogramming by rocaglates, cap-dependent initiation disruptors that include zotatifin. We find effects to be far more complex than simple “translational inhibition” as currently defined. Translatome analysis by TMT-pSILAC (tandem mass tag-pulse stable isotope labeling with amino acids in cell culture mass spectrometry) reveals myriad upregulated proteins that drive hitherto unrecognized cytotoxic mechanisms, including GEF-H1-mediated anti-survival RHOA/JNK activation. Surprisingly, these responses are not replicated by eIF4A silencing, indicating a broader translational adaptation than currently understood. Translation machinery analysis by MATRIX (mass spectrometry analysis of active translation factors using ribosome density fractionation and isotopic labeling experiments) identifies rocaglate-specific dependence on specific translation factors including eEF1ε1 that drive translatome remodeling. Our proteome-level interrogation reveals that the complete cellular response to these historical “translation inhibitors” is mediated by comprehensive translational landscape remodeling.

Original languageEnglish (US)
Article number109806
JournalCell Reports
Volume37
Issue number2
DOIs
StatePublished - Oct 12 2021

Keywords

  • DDX17
  • GEF-H1
  • JNK
  • RHOA
  • eEF1ε1
  • eIF4A
  • rocaglate
  • silvestrol
  • translation
  • zotatifin

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Fingerprint

Dive into the research topics of 'Proteomics reveal cap-dependent translation inhibitors remodel the translation machinery and translatome'. Together they form a unique fingerprint.

Cite this