A substrate binding model for the KEOPS tRNA modifying complex

Nat Commun. 2020 Dec 4;11(1):6233. doi: 10.1038/s41467-020-19990-5.

Abstract

The KEOPS complex, which is conserved across archaea and eukaryotes, is composed of four core subunits; Pcc1, Kae1, Bud32 and Cgi121. KEOPS is crucial for the fitness of all organisms examined. In humans, pathogenic mutations in KEOPS genes lead to Galloway-Mowat syndrome, an autosomal-recessive disease causing childhood lethality. Kae1 catalyzes the universal and essential tRNA modification N6-threonylcarbamoyl adenosine, but the precise roles of all other KEOPS subunits remain an enigma. Here we show using structure-guided studies that Cgi121 recruits tRNA to KEOPS by binding to its 3' CCA tail. A composite model of KEOPS bound to tRNA reveals that all KEOPS subunits form an extended tRNA-binding surface that we have validated in vitro and in vivo to mediate the interaction with the tRNA substrate and its modification. These findings provide a framework for understanding the inner workings of KEOPS and delineate why all KEOPS subunits are essential.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Archaeal Proteins / chemistry*
  • Archaeal Proteins / genetics
  • Archaeal Proteins / metabolism
  • Crystallography, X-Ray
  • Methanocaldococcus / genetics
  • Methanocaldococcus / metabolism*
  • Models, Molecular
  • Multiprotein Complexes / chemistry*
  • Multiprotein Complexes / genetics
  • Multiprotein Complexes / metabolism
  • Nucleic Acid Conformation
  • Protein Binding
  • Protein Domains
  • RNA, Transfer / chemistry*
  • RNA, Transfer / genetics
  • RNA, Transfer / metabolism
  • RNA, Transfer, Lys / chemistry
  • RNA, Transfer, Lys / genetics
  • RNA, Transfer, Lys / metabolism

Substances

  • Archaeal Proteins
  • Multiprotein Complexes
  • RNA, Transfer, Lys
  • RNA, Transfer