Activation of the Legionella pneumophila LegK7 Effector Kinase by the Host MOB1 Protein

J Mol Biol. 2021 Feb 5;433(3):166746. doi: 10.1016/j.jmb.2020.166746. Epub 2020 Dec 10.

Abstract

Legionella pneumophila infects alveolar macrophages and can cause life-threatening pneumonia in humans. Upon internalization into the host cell, L. pneumophila injects numerous effector proteins into the host cytoplasm as a part of its pathogenesis. LegK7 is an effector kinase of L. pneumophila that functionally mimics the eukaryotic Mst kinase and phosphorylates the host MOB1 protein to exploit the Hippo pathway. To elucidate the LegK7 activation mechanism, we determined the apo structure of LegK7 in an inactive form and performed a comparative analysis of LegK7 structures. LegK7 is a non-RD kinase that contains an activation segment that is ordered, irrespective of stimulation, through a unique β-hairpin-containing segment, and it does not require phosphorylation of the activation segment for activation. Instead, bacterial LegK7 becomes an active kinase via its heterologous molecular interaction with the host MOB1 protein. MOB1 binding triggers reorientation of the two lobes of the kinase domain, as well as a structural change in the interlobe hinge region in LegK7, consequently reshaping the LegK7 structure into an ATP binding-compatible closed conformation. Furthermore, we reveal that LegK7 is an atypical kinase that contains an N-terminal capping domain and a hydrophilic interlobe linker motif, which play key roles in the MOB1-induced activation of LegK7.

Keywords: LegK7; Legionella pneumophila; MOB1; protein kinase; structural rearrangement.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chemokine CXCL10 / chemistry
  • Chemokine CXCL10 / genetics
  • Chemokine CXCL10 / metabolism*
  • Enzyme Activation
  • Host-Pathogen Interactions*
  • Humans
  • Legionella pneumophila / enzymology*
  • Legionnaires' Disease / metabolism*
  • Legionnaires' Disease / microbiology*
  • Molecular Docking Simulation
  • Molecular Dynamics Simulation
  • Phosphorylation
  • Protein Binding
  • Protein Conformation
  • Protein Interaction Domains and Motifs
  • Protein Kinases / chemistry
  • Protein Kinases / metabolism*
  • Spectrum Analysis
  • Structure-Activity Relationship

Substances

  • CXCL10 protein, human
  • Chemokine CXCL10
  • Protein Kinases