Proteomics

Dataset Information

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Identification of the endogenous host targets of the Salmonella effectors SSeK1 and 3


ABSTRACT: Strains of Salmonella utilise two distinct type three secretion systems to deliver effector proteins directly into host cells. The Salmonella effectors SseK1 and SseK3 are arginine glycosyltransferases that modify mammalian death domain containing proteins with N-acetyl glucosamine (GlcNAc) when overexpressed ectopically or as recombinant protein fusions. Here, we combined Arg-GlcNAc glycopeptide immunoprecipitation and mass spectrometry to identify host proteins GlcNAcylated by endogenous levels of SseK1 and SseK3 during Salmonella infection. We observed that SseK1 modified the mammalian signaling protein TRADD, but not FADD as previously reported. Overexpression of SseK1 greatly broadened substrate specificity, while ectopic co-expression of SseK1 and TRADD increased the range of modified arginine residues within the death domain of TRADD. In contrast, endogenous levels of SseK3 resulted in modification of the death domains of receptors of the mammalian TNF superfamily, TNFR1 and TRAILR, at residues Arg376 and Arg293 respectively. Structural studies on SseK3 showed that the enzyme displays a classic GT-A glycosyltransferase fold and binds UDP-GlcNAc in a narrow and deep cleft with the GlcNAc facing the surface. Together our data suggests that Salmonellae carrying sseK1 and sseK3 employ the glycosyltransferase effectors to antagonise different components of death receptor signaling.

INSTRUMENT(S): Orbitrap Fusion Lumos, LTQ Orbitrap Elite

ORGANISM(S): Homo Sapiens (human) Salmonella Enterica Subsp. Enterica Serovar Typhimurium Str. Sl1344 Mus Musculus (mouse)

TISSUE(S): Macrophage

SUBMITTER: Nichollas Scott  

LAB HEAD: Elizabeth L Hartland

PROVIDER: PXD010769 | Pride | 2019-03-27

REPOSITORIES: Pride

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Publications

<i>Salmonella</i> Effectors SseK1 and SseK3 Target Death Domain Proteins in the TNF and TRAIL Signaling Pathways.

Newson Joshua P M JPM   Scott Nichollas E NE   Yeuk Wah Chung Ivy I   Wong Fok Lung Tania T   Giogha Cristina C   Gan Jiyao J   Wang Nancy N   Strugnell Richard A RA   Brown Nathaniel F NF   Cygler Miroslaw M   Pearson Jaclyn S JS   Hartland Elizabeth L EL  

Molecular & cellular proteomics : MCP 20190322 6


Strains of <i>Salmonella</i> utilize two distinct type three secretion systems to deliver effector proteins directly into host cells. The <i>Salmonella</i> effectors SseK1 and SseK3 are arginine glycosyltransferases that modify mammalian death domain containing proteins with <i>N</i>-acetyl glucosamine (GlcNAc) when overexpressed ectopically or as recombinant protein fusions. Here, we combined Arg-GlcNAc glycopeptide immunoprecipitation and mass spectrometry to identify host proteins GlcNAcylate  ...[more]

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