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TitleStructural basis for the toxicity of Legionella pneumophila effector SidH.
Journal, issue, pagesNat Commun, Vol. 14, Issue 1, Page 7068, Year 2023
Publish dateNov 3, 2023
AuthorsRahul Sharma / Michael Adams / Simonne Griffith-Jones / Tobias Sahr / Laura Gomez-Valero / Felix Weis / Michael Hons / Sarah Gharbi / Rayene Berkane / Alexandra Stolz / Carmen Buchrieser / Sagar Bhogaraju /
PubMed AbstractLegionella pneumophila (LP) secretes more than 300 effectors into the host cytosol to facilitate intracellular replication. One of these effectors, SidH, 253 kDa in size with no sequence similarity ...Legionella pneumophila (LP) secretes more than 300 effectors into the host cytosol to facilitate intracellular replication. One of these effectors, SidH, 253 kDa in size with no sequence similarity to proteins of known function is toxic when overexpressed in host cells. SidH is regulated by the LP metaeffector LubX which targets SidH for degradation in a temporal manner during LP infection. The mechanism underlying the toxicity of SidH and its role in LP infection are unknown. Here, we determined the cryo-EM structure of SidH at 2.7 Å revealing a unique alpha helical arrangement with no overall similarity to known protein structures. Surprisingly, purified SidH came bound to a E. coli EF-Tu/t-RNA/GTP ternary complex which could be modeled into the cryo-EM density. Mutation of residues disrupting the SidH-tRNA interface and SidH-EF-Tu interface abolish the toxicity of overexpressed SidH in human cells, a phenotype confirmed in infection of Acanthamoeba castellani. We also present the cryo-EM structure of SidH in complex with a U-box domain containing ubiquitin ligase LubX delineating the mechanism of regulation of SidH. Our data provide the basis for the toxicity of SidH and into its regulation by the metaeffector LubX.
External linksNat Commun / PubMed:37923743 / PubMed Central
MethodsEM (single particle)
Resolution2.7 - 3.1 Å
Structure data

18383

8qfs

EMDB-18383, PDB-8qfs:
Cryo-EM structure of SidH from Legionella pneumophila
Method: EM (single particle) / Resolution: 2.7 Å

18407

8qhc

EMDB-18407, PDB-8qhc:
Cryo-EM structure of SidH from Legionella pneumophila in complex with LubX
Method: EM (single particle) / Resolution: 3.1 Å

Chemicals

ChemComp-MG:
Unknown entry

ChemComp-GTP:
GUANOSINE-5'-TRIPHOSPHATE / GTP, energy-carrying molecule*YM / Guanosine triphosphate

Source
  • legionella pneumophila (bacteria)
  • escherichia coli 'bl21-gold(de3)plyss ag' (bacteria)
KeywordsRNA BINDING PROTEIN / SidH / Legionella / tRNA-binding / EF-Tu-binding

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