Arg-GlcNAcylation on TRADD by NleB and SseK1 Is Crucial for Bacterial Pathogenesis

Xue, Jian; Hu, Shufan; Huang, Yuxuan; Zhang, Qi; Yi, Xueying; Xing, Puyuan; Li, Shan

doi:10.3389/fcell.2020.00641

Cited by 18 publications

(11 citation statements)

References 47 publications

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“…Salmonella enterica serovar typhimurium can also modify some DD host proteins through arginine-GlcNAcylation to block death receptor-mediated proinflammatory responses. T3SEs SseK3 and SseK1 are specialized to the modification of tumor necrosis factor receptor type 1 (TNFR1) and TNFR1-associated DD (TRADD) protein, respectively [ 71 ]. SseK1, SseK3 and NleB structures revealed a very high degree of structural similarity [ 70 ].…”

Section: The Diverse Roles Of α-Helical Domains In Animal T3s Effectorsmentioning

confidence: 99%

α-Helices in the Type III Secretion Effectors: A Prevalent Feature with Versatile Roles

Gazi

Kokkinidis

Fadouloglou

2021

IJMS

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Type III Secretion Systems (T3SSs) are multicomponent nanomachines located at the cell envelope of Gram-negative bacteria. Their main function is to transport bacterial proteins either extracellularly or directly into the eukaryotic host cell cytoplasm. Type III Secretion effectors (T3SEs), latest to be secreted T3S substrates, are destined to act at the eukaryotic host cell cytoplasm and occasionally at the nucleus, hijacking cellular processes through mimicking eukaryotic proteins. A broad range of functions is attributed to T3SEs, ranging from the manipulation of the host cell’s metabolism for the benefit of the bacterium to bypassing the host’s defense mechanisms. To perform this broad range of manipulations, T3SEs have evolved numerous novel folds that are compatible with some basic requirements: they should be able to easily unfold, pass through the narrow T3SS channel, and refold to an active form when on the other side. In this review, the various folds of T3SEs are presented with the emphasis placed on the functional and structural importance of α-helices and helical domains.

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Section: The Diverse Roles Of α-Helical Domains In Animal T3s Effectorsmentioning

confidence: 99%

α-Helices in the Type III Secretion Effectors: A Prevalent Feature with Versatile Roles

Gazi

Kokkinidis

Fadouloglou

2021

IJMS

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“…Multiple host protein substrates for the NleB/SseK orthologs have been described. TRADD, FADD, RIPK1, HIF-1α, and GAPDH are reported to be the host targets for EPEC NleB ( Gao et al, 2013 ; Li et al, 2013 ; El Qaidi et al, 2017 ; Gunster et al, 2017 ; Scott et al, 2017 ; Ding et al, 2019 ; Newson et al, 2019 ; Xue et al, 2020a ; Garcia-Garcia et al, 2021 ). We and other groups have reported TRADD is a in vivo substrate of NleB and SseK1.…”

Section: Introductionmentioning

confidence: 99%

“…We and other groups have reported TRADD is a in vivo substrate of NleB and SseK1. Arg-GlcNAcylation on TRADD by NleB and SseK1 is crucial for bacterial pathogenesis ( Newson et al, 2019 ; Xue et al, 2020a ). SseK3 was also found to affect bacterial virulence by modifying signaling receptors TNFR1, TRAILR, and phylogenetically related Rab GTPases ( Newson et al, 2019 ; Gan et al, 2020 ; Meng et al, 2020 ; Xue et al, 2020a ).…”

Section: Introductionmentioning

confidence: 99%

“…Arg-GlcNAcylation on TRADD by NleB and SseK1 is crucial for bacterial pathogenesis ( Newson et al, 2019 ; Xue et al, 2020a ). SseK3 was also found to affect bacterial virulence by modifying signaling receptors TNFR1, TRAILR, and phylogenetically related Rab GTPases ( Newson et al, 2019 ; Gan et al, 2020 ; Meng et al, 2020 ; Xue et al, 2020a ). In addition to glycosylating host proteins, NleB, SseK1, and SseK3 could also glycosylate themselves ( Park et al, 2018 ; Newson et al, 2019 ; Xue et al, 2020b ; Garcia-Garcia et al, 2021 ).…”

Section: Introductionmentioning

confidence: 99%

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Arginine GlcNAcylation and Activity Regulation of PhoP by a Type III Secretion System Effector in Salmonella

et al. 2022

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Salmonella type III secretion system (T3SS) effector SseK3 is a glycosyltransferase delivered directly into the host cells to modify host protein substrates, thus manipulating host cellular signal transduction. Here, we identify and characterize the Arg-GlcNAcylation activity of SseK3 inside bacterial cells. Combining Arg-GlcNAc protein immunoprecipitation and mass spectrometry, we found that 60 bacterial proteins were GlcNAcylated during Salmonella infection, especially the two-component signal transduction system regulatory protein PhoP. Moreover, the Arg-GlcNAcylation of PhoP by SseK3 was detected in vivo and in vitro, and four arginine residues, Arg65, Arg66, Arg118, and Arg215 were identified as the GlcNAcylation sites. Site-directed mutagenesis showed that the PhoP R215A change significantly reduced the DNA-binding ability and arginine to alanine change at all four sites (PhoP 4RA) completely eliminated the DNA-binding ability, suggesting that Arg215 is essential for the DNA-binding activity of PhoP and GlcNAcylation of PhoP affects this activity. Additionally, GlcNAcylation of PhoP negatively regulated the activity of PhoP and decreased the expression of its downstream genes. Overall, our work provides an example of the intra-bacterial activities of the T3SS effectors and increases our understanding of endogenous Arg-GlcNAcylation.

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“…Homologues of NleB1 are found in Citrobacter rodentium (termed NleB) and Salmonella species (termed SseK1, SseK2, SseK3) [6][7][8]. These homologues have well-characterized roles in inhibition of death receptor signalling [4,5,[9][10][11][12][13][14][15] and some also block small Rab GTPase function [16,17]. However, EHEC and EPEC strains also contain the NleB1 paralogue NleB2, which is less well-characterized.…”

Section: Introductionmentioning

confidence: 99%

NleB2 from enteropathogenic Escherichia coli is a novel arginine-glucose transferase effector

et al. 2021

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During infection, enteropathogenic Escherichia coli (EPEC) and enterohaemorrhagic E. coli (EHEC) directly manipulate various aspects of host cell function through the translocation of type III secretion system (T3SS) effector proteins directly into the host cell. Many T3SS effector proteins are enzymes that mediate post-translational modifications of host proteins, such as the glycosyltransferase NleB1, which transfers a single N-acetylglucosamine (GlcNAc) to arginine residues, creating an Arg-GlcNAc linkage. NleB1 glycosylates death-domain containing proteins including FADD, TRADD and RIPK1 to block host cell death. The NleB1 paralogue, NleB2, is found in many EPEC and EHEC strains but to date its enzymatic activity has not been described. Using in vitro glycosylation assays combined with mass spectrometry, we found that NleB2 can utilize multiple sugar donors including UDP-glucose, UDP-GlcNAc and UDP-galactose during glycosylation of the death domain protein, RIPK1. Sugar donor competition assays demonstrated that UDP-glucose was the preferred substrate of NleB2 and peptide sequencing identified the glycosylation site within RIPK1 as Arg603, indicating that NleB2 catalyses arginine glucosylation. We also confirmed that NleB2 catalysed arginine-hexose modification of Flag-RIPK1 during infection of HEK293T cells with EPEC E2348/69. Using site-directed mutagenesis and in vitro glycosylation assays, we identified that residue Ser252 in NleB2 contributes to the specificity of this distinct catalytic activity. Substitution of Ser252 in NleB2 to Gly, or substitution of the corresponding Gly255 in NleB1 to Ser switches sugar donor preference between UDP-GlcNAc and UDP-glucose. However, this switch did not affect the ability of the NleB variants to inhibit inflammatory or cell death signalling during HeLa cell transfection or EPEC infection. NleB2 is thus the first identified bacterial Arg-glucose transferase that, similar to the NleB1 Arg-GlcNAc transferase, inhibits host protein function by arginine glycosylation.

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Arg-GlcNAcylation on TRADD by NleB and SseK1 Is Crucial for Bacterial Pathogenesis

Cited by 18 publications

References 47 publications

α-Helices in the Type III Secretion Effectors: A Prevalent Feature with Versatile Roles

α-Helices in the Type III Secretion Effectors: A Prevalent Feature with Versatile Roles

Arginine GlcNAcylation and Activity Regulation of PhoP by a Type III Secretion System Effector in Salmonella

NleB2 from enteropathogenic Escherichia coli is a novel arginine-glucose transferase effector

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