Spatiotemporal proteomics uncovers cathepsin-dependent macrophage cell death during Salmonella infection.
Animals
Cathepsins
/ drug effects
Cell Death
/ drug effects
Cystatin B
/ antagonists & inhibitors
Inflammasomes
/ metabolism
Intracellular Signaling Peptides and Proteins
/ metabolism
Lipopolysaccharides
/ metabolism
Lysosomes
/ metabolism
Macrophages
/ metabolism
Mice
Peptide Hydrolases
/ metabolism
Phosphate-Binding Proteins
/ metabolism
Proteome
Proteomics
RAW 264.7 Cells
Salmonella Infections
/ metabolism
Salmonella typhimurium
/ metabolism
Journal
Nature microbiology
ISSN: 2058-5276
Titre abrégé: Nat Microbiol
Pays: England
ID NLM: 101674869
Informations de publication
Date de publication:
09 2020
09 2020
Historique:
received:
08
11
2018
accepted:
06
05
2020
pubmed:
10
6
2020
medline:
18
11
2020
entrez:
10
6
2020
Statut:
ppublish
Résumé
The interplay between host and pathogen relies heavily on rapid protein synthesis and accurate protein targeting to ensure pathogen destruction. To gain insight into this dynamic interface, we combined Click chemistry with pulsed stable isotope labelling of amino acids in cell culture to quantify the host proteome response during macrophage infection with the intracellular bacterial pathogen Salmonella enterica Typhimurium. We monitored newly synthesized proteins across different host cell compartments and infection stages. Within this rich resource, we detected aberrant trafficking of lysosomal proteases to the extracellular space and the nucleus. We verified that active cathepsins re-traffic to the nucleus and that these are linked to cell death. Pharmacological cathepsin inhibition and nuclear targeting of a cellular cathepsin inhibitor (stefin B) suppressed S. enterica Typhimurium-induced cell death. We demonstrate that cathepsin activity is required for pyroptotic cell death via the non-canonical inflammasome, and that lipopolysaccharide transfection into the host cytoplasm is sufficient to trigger active cathepsin accumulation in the host nucleus and cathepsin-dependent cell death. Finally, cathepsin inhibition reduced gasdermin D expression, thus revealing an unexpected role for cathepsin activity in non-canonical inflammasome regulation. Overall, our study illustrates how resolution of host proteome dynamics during infection can drive the discovery of biological mechanisms at the host-microbe interface.
Identifiants
pubmed: 32514074
doi: 10.1038/s41564-020-0736-7
pii: 10.1038/s41564-020-0736-7
pmc: PMC7610801
mid: EMS118378
doi:
Substances chimiques
Gsdmd protein, mouse
0
Inflammasomes
0
Intracellular Signaling Peptides and Proteins
0
Lipopolysaccharides
0
Phosphate-Binding Proteins
0
Proteome
0
Cystatin B
88844-95-5
Cathepsins
EC 3.4.-
Peptide Hydrolases
EC 3.4.-
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1119-1133Références
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