6XNJ

Crystal structure of the PDZ domain of human GOPC in complex with a peptide of E. coli O157:H7 str. Sakai effector NleG8

Summary for 6XNJ

• Entry DOI: 10.2210/pdb6xnj/pdb
• Descriptor: Golgi-associated PDZ and coiled-coil motif-containing protein, NleG8 peptide, SULFATE ION, ... (4 entities in total)
• Functional Keywords: ubiquitination, effectors, structural genomics, center for structural genomics of infectious diseases, csgid, protein binding
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 2
• Total formula weight: 11050.58

Authors

Stogios, P.J.,Skarina, T.,Popov, G.,Chang, C.,Savchenko, A.,Joachimiak, A.,Satchell, K.J.F.,Center for Structural Genomics of Infectious Diseases (CSGID) (deposition date: 2020-07-03, release date: 2020-08-12, Last modification date: 2026-08-19)

Primary citation

Popov, G.,Fiebig-Comyn, A.,Syriste, L.,Little, D.J.,Skarina, T.,Stogios, P.J.,Birstonas, S.,Coombes, B.K.,Savchenko, A.
Distinct Molecular Features of NleG Type 3 Secreted Effectors Allow for Different Roles during Citrobacter rodentium Infection in Mice.
Infect.Immun., 91:e0050522-e0050522, 2023

PubMed Abstract: The NleGs are the largest family of type 3 secreted effectors in attaching and effacing (A/E) pathogens, such as enterohemorrhagic Escherichia coli (EHEC), enteropathogenic E. coli, and Citrobacter rodentium. NleG effectors contain a conserved C-terminal U-box domain acting as a ubiquitin protein ligase and target host proteins via a variable N-terminal portion. The specific roles of these effectors during infection remain uncertain. Here, we demonstrate that the three NleG effectors-NleG1, NleG7, and NleG8-encoded by C. rodentium DBS100 play distinct roles during infection in mice. Using individual knockout strains, we show that NleG7 contributes to bacterial survival during enteric infection while NleG1 promotes the expression of diarrheal symptoms and NleG8 contributes to accelerated lethality in susceptible mice. Furthermore, the NleG8 effector contains a C-terminal PDZ domain binding motif that enables interaction with the host protein GOPC. Both the PDZ domain binding motif and the ability to engage with host ubiquitination machinery via the intact U-box domain proved to be necessary for NleG8 function, contributing to the observed phenotype during infection. We also establish that the PTZ binding motif in the EHEC NleG8 (NleG8) effector, which shares 60% identity with NleG8, is engaged in interactions with human GOPC. The crystal structure of the NleG8 C-terminal peptide in complex with the GOPC PDZ domain, determined to 1.85 Å, revealed a conserved interaction mode similar to that observed between GOPC and eukaryotic PDZ domain binding motifs. Despite these common features, does not complement the Δ phenotype during infection, revealing functional diversification between these NleG effectors.

PubMed: 36511702
DOI: 10.1128/iai.00505-22

Experimental method

X-RAY DIFFRACTION (1.85 Å)