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TitleMolecular basis of A. thaliana KEOPS complex in biosynthesizing tRNA t6A.
Journal, issue, pagesNucleic Acids Res, Year 2024
Publish dateMar 13, 2024
AuthorsXinxing Zheng / Chenchen Su / Lei Duan / Mengqi Jin / Yongtao Sun / Li Zhu / Wenhua Zhang /
PubMed AbstractIn archaea and eukaryotes, the evolutionarily conserved KEOPS is composed of four core subunits-Kae1, Bud32, Cgi121 and Pcc1, and a fifth Gon7/Pcc2 that is found in fungi and metazoa. KEOPS ...In archaea and eukaryotes, the evolutionarily conserved KEOPS is composed of four core subunits-Kae1, Bud32, Cgi121 and Pcc1, and a fifth Gon7/Pcc2 that is found in fungi and metazoa. KEOPS cooperates with Sua5/YRDC to catalyze the biosynthesis of tRNA N6-threonylcarbamoyladenosine (t6A), an essential modification needed for fitness of cellular organisms. Biochemical and structural characterizations of KEOPSs from archaea, yeast and humans have determined a t6A-catalytic role for Kae1 and auxiliary roles for other subunits. However, the precise molecular workings of KEOPSs still remain poorly understood. Here, we investigated the biochemical functions of A. thaliana KEOPS and determined a cryo-EM structure of A. thaliana KEOPS dimer. We show that A. thaliana KEOPS is composed of KAE1, BUD32, CGI121 and PCC1, which adopts a conserved overall arrangement. PCC1 dimerization leads to a KEOPS dimer that is needed for an active t6A-catalytic KEOPS-tRNA assembly. BUD32 participates in direct binding of tRNA to KEOPS and modulates the t6A-catalytic activity of KEOPS via its C-terminal tail and ATP to ADP hydrolysis. CGI121 promotes the binding of tRNA to KEOPS and potentiates the t6A-catalytic activity of KEOPS. These data and findings provide insights into mechanistic understanding of KEOPS machineries.
External linksNucleic Acids Res / PubMed:38477398
MethodsEM (single particle)
Resolution3.7 Å
Structure data

36808

8k20

EMDB-36808, PDB-8k20:
Cryo-EM structure of KEOPS complex from Arabidopsis thaliana
Method: EM (single particle) / Resolution: 3.7 Å

Chemicals

ChemComp-FE:
Unknown entry / Iron

Source
  • arabidopsis thaliana (thale cress)
KeywordsTRANSFERASE / KEOPS complex / tRNA t6A synthase / RNA-binding protein

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