Mutagenesis and Functional Analysis of the Bacterial Arginine Glycosyltransferase Effector NleB1 from Enteropathogenic Escherichia coli

Infection and Immunity
Tania Wong Fok LungElizabeth L Hartland

Abstract

Enteropathogenic Escherichia coli (EPEC) interferes with host cell signaling by injecting virulence effector proteins into enterocytes via a type III secretion system (T3SS). NleB1 is a novel T3SS glycosyltransferase effector from EPEC that transfers a single N-acetylglucosamine (GlcNAc) moiety in an N-glycosidic linkage to Arg(117) of the Fas-associated death domain protein (FADD). GlcNAcylation of FADD prevents the assembly of the canonical death-inducing signaling complex and inhibits Fas ligand (FasL)-induced cell death. Apart from the DXD catalytic motif of NleB1, little is known about other functional sites in the enzyme. In the present study, members of a library of 22 random transposon-based, in-frame, linker insertion mutants of NleB1 were tested for their ability to block caspase-8 activation in response to FasL during EPEC infection. Immunoblot analysis of caspase-8 cleavage showed that 17 mutant derivatives of NleB1, including the catalytic DXD mutant, did not inhibit caspase-8 activation. Regions of interest around the insertion sites with multiple or single amino acid substitutions were examined further. Coimmunoprecipitation studies of 34 site-directed mutants showed that the NleB1 derivatives with the E253A, Y21...Continue Reading

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Nov 29, 2016·Annual Review of Genetics·Jaclyn S PearsonElizabeth L Hartland
Jan 18, 2018·Infection and Immunity·Maarten F de Jong, Neal M Alto
Feb 17, 2018·The Journal of Biological Chemistry·Diego EspositoKatrin Rittinger
Sep 2, 2017·The Journal of Biological Chemistry·Nichollas E ScottElizabeth L Hartland
Feb 1, 2017·Infection and Immunity·Georgina L PollockElizabeth L Hartland
Mar 25, 2019·Molecular & Cellular Proteomics : MCP·Joshua P M NewsonElizabeth L Hartland
Mar 7, 2020·Microorganisms·Juan Luis Araujo-GarridoFrancisco Ramos-Morales
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May 16, 2020·Frontiers in Cellular and Infection Microbiology·Xing PanShan Li
Jun 1, 2019·Molecular & Cellular Proteomics : MCP·JoshuaP M NewsonElizabeth L Hartland
Mar 25, 2021·Current Opinion in Structural Biology·Eunhee Koh, Hyun-Soo Cho
Jun 17, 2021·PLoS Pathogens·Cristina GioghaElizabeth L Hartland

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