title: The Shigella type three secretion system effector OspG directly and specifically binds to host ubiquitin for activation.
abstract: The genus Shigella infects human gut epithelial cells to cause diarrhea and gastrointestinal disorders. Like many other Gram-negative bacterial pathogens, the virulence of Shigella spp. relies on a conserved type three secretion system that delivers a handful of effector proteins into host cells to manipulate various host cell physiology. However, many of the Shigella type III effectors remain functionally uncharacterized. Here we observe that OspG, one of the Shigella effectors, interacted with ubiquitin conjugates and poly-ubiquitin chains of either K48 or K63 linkage in eukaryotic host cells. Purified OspG protein formed a stable complex with ubiquitin but showed no interactions with other ubiquitin-like proteins. OspG binding to ubiquitin required the carboxyl terminal helical region in OspG and the canonical I44-centered hydrophobic surface in ubiquitin. OspG and OspG-homologous effectors, NleH1/2 from enteropathogenic E coli (EPEC), contain sub-domains I-VII of eukaryotic serine/threonine kinase. GST-tagged OspG and NleH1/2 could undergo autophosphorylation, the former of which was significantly stimulated by ubiquitin binding. Ubiquitin binding was also required for OspG functioning in attenuating host NF-κB signaling. Our data illustrate a new mechanism that bacterial pathogen like Shigella exploits ubiquitin binding to activate its secreted virulence effector for its functioning in host eukaryotic cells.
authors.affiliation: College of Life Sciences, Beijing Normal University, Beijing, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhou
authors.forename: Na
authors.initials: N
authors.lastname: Dong
authors.forename: Liyan
authors.initials: L
authors.lastname: Hu
authors.forename: Feng
authors.initials: F
authors.lastname: Shao
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D048057:Mitogen-Activated Protein Kinase 10 / Q000737:chemistry; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism; D011485:Protein Binding; D017386:Sequence Homology, Amino Acid; D012760:Shigella / Q000378:metabolism / Q000502:physiology*; D015398:Signal Transduction; D025801:Ubiquitin / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D016328:NF-kappa B; D025801:Ubiquitin; D048057:Mitogen-Activated Protein Kinase 10