Structural and mechanistic basis for protein glutamylation by the kinase fold

Adam Osinski, Miles H. Black, Krzysztof Pawłowski, Zhe Chen, Yang Li, Vincent S. Tagliabracci

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

The kinase domain transfers phosphate from ATP to substrates. However, the Legionella effector SidJ adopts a kinase fold, yet catalyzes calmodulin (CaM)-dependent glutamylation to inactivate the SidE ubiquitin ligases. The structural and mechanistic basis in which the kinase domain catalyzes protein glutamylation is unknown. Here we present cryo-EM reconstructions of SidJ:CaM:SidE reaction intermediate complexes. We show that the kinase-like active site of SidJ adenylates an active-site Glu in SidE, resulting in the formation of a stable reaction intermediate complex. An insertion in the catalytic loop of the kinase domain positions the donor Glu near the acyl-adenylate for peptide bond formation. Our structural analysis led us to discover that the SidJ paralog SdjA is a glutamylase that differentially regulates the SidE ligases during Legionella infection. Our results uncover the structural and mechanistic basis in which the kinase fold catalyzes non-ribosomal amino acid ligations and reveal an unappreciated level of SidE-family regulation.

Original languageEnglish (US)
Pages (from-to)4527-4539.e8
JournalMolecular cell
Volume81
Issue number21
DOIs
StatePublished - Nov 4 2021

Keywords

  • effectors
  • glutamylation
  • Legionella
  • pseudokinase
  • SdeA
  • SdeB
  • SdeC
  • SdjA
  • SidE
  • SidJ

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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