Demonstration of translation elongation factor 3 activity from a non-fungal species, Phytophthora infestans

PLoS One. 2018 Jan 4;13(1):e0190524. doi: 10.1371/journal.pone.0190524. eCollection 2018.

Abstract

In most eukaryotic organisms, translation elongation requires two highly conserved elongation factors eEF1A and eEF2. Fungal systems are unique in requiring a third factor, the eukaryotic Elongation Factor 3 (eEF3). For decades, eEF3, a ribosome-dependent ATPase, was considered "fungal-specific", however, recent bioinformatics analysis indicates it may be more widely distributed among other unicellular eukaryotes. In order to determine whether divergent eEF3-like proteins from other eukaryotic organisms can provide the essential functions of eEF3 in budding yeast, the eEF3-like proteins from Schizosaccharomyes pombe and an oomycete, Phytophthora infestans, were cloned and expressed in Saccharomyces cerevisiae. Plasmid shuffling experiments showed that both S. pombe and P. infestans eEF3 can support the growth of S. cerevisiae in the absence of endogenous budding yeast eEF3. Consistent with its ability to provide the essential functions of eEF3, P. infestans eEF3 possessed ribosome-dependent ATPase activity. Yeast cells expressing P. infestans eEF3 displayed reduced protein synthesis due to defects in translation elongation/termination. Identification of eEF3 in divergent species will advance understanding of its function and the ribosome specific determinants that lead to its requirement as well as contribute to the identification of functional domains of eEF3 for potential drug discovery.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Peptide Chain Elongation, Translational
  • Peptide Elongation Factors / classification
  • Peptide Elongation Factors / metabolism*
  • Phylogeny
  • Phytophthora infestans / metabolism*

Substances

  • Peptide Elongation Factors

Grants and funding

This work was supported by the New Jersey Health Foundation (njhealthfoundation.org) grant number PC 11-17 to T.G.K., and by Novo Nordisk (novonordisk-us.com) grant number 10307 to S.D. The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.