Structural and mechanistic basis for protein glutamylation by the kinase fold

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

doi:10.1016/j.molcel.2021.08.007

Title: Structural and mechanistic basis for protein glutamylation by the kinase fold

Journal Article · 16 August 2021 · Molecular Cell

DOI: https://doi.org/10.1016/j.molcel.2021.08.007 · OSTI ID:1981712

^[1];

^[2]; Chen, Zhe ^[1]; Li, Yang ^[1]; Tagliabracci, Vincent S. ^[1]

Univ. of Texas Southwestern Medical Center, Dallas, TX (United States)
Univ. of Texas Southwestern Medical Center, Dallas, TX (United States); Warsaw Univ. of Life Sciences (Poland)

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, in this work, 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.

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Research Organization:: Pacific Northwest National Laboratory (PNNL), Richland, WA (United States). Environmental Molecular Sciences Laboratory (EMSL)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: AC05-76RL01830

OSTI ID:: 1981712

Journal Information:: Molecular Cell, Journal Name: Molecular Cell Journal Issue: 21 Vol. 81; ISSN 1097-2765

Publisher:: Cell Press - ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
Legionella
SdeA
SdeB
SdeC
SdjA
SidE
SidJ
effectors
glutamylation
pseudokinase

Title: Structural and mechanistic basis for protein glutamylation by the kinase fold

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