title: Systematic analysis of bacterial effector-postsynaptic density 95/disc large/zonula occludens-1 (PDZ) domain interactions demonstrates Shigella OspE protein promotes protein kinase C activation via PDLIM proteins.
abstract: Diseases caused by many Gram-negative bacterial pathogens depend on the activities of bacterial effector proteins that are delivered into eukaryotic cells via specialized secretion systems. Effector protein function largely depends on specific subcellular targeting and specific interactions with cellular ligands. PDZ domains are common domains that serve to provide specificity in protein-protein interactions in eukaryotic systems. We show that putative PDZ-binding motifs are significantly enriched among effector proteins delivered into mammalian cells by certain bacterial pathogens. We use PDZ domain microarrays to identify candidate interaction partners of the Shigella flexneri effector proteins OspE1 and OspE2, which contain putative PDZ-binding motifs. We demonstrate in vitro and in cells that OspE proteins interact with PDLIM7, a member of the PDLIM family of proteins, which contain a PDZ domain and one or more LIM domains, protein interaction domains that participate in a wide variety of functions, including activation of isoforms of protein kinase C (PKC). We demonstrate that activation of PKC during S. flexneri infection is attenuated in the absence of PDLIM7 or OspE proteins and that the OspE PDZ-binding motif is required for wild-type levels of PKC activation. These results are consistent with a model in which binding of OspE to PDLIM7 during infection regulates the activity of PKC isoforms that bind to the PDLIM7 LIM domain.
authors.affiliation: From the Division of Infectious Diseases, Massachusetts General Hospital, Cambridge, Massachusetts 02139.
authors.forename: Chae-ryun
authors.initials: CR
authors.lastname: Yi
authors.affiliation: the Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts 02138, and.
authors.forename: John E
authors.initials: JE
authors.lastname: Allen
authors.affiliation: From the Division of Infectious Diseases, Massachusetts General Hospital, Cambridge, Massachusetts 02139, Departments of Microbiology and Immunobiology and.
authors.forename: Brian
authors.initials: B
authors.lastname: Russo
authors.affiliation: the Program in Immunology and.
authors.forename: Soo Young
authors.initials: SY
authors.lastname: Lee
authors.affiliation: Departments of Microbiology and Immunobiology and.
authors.forename: Jason E
authors.initials: JE
authors.lastname: Heindl
authors.affiliation: From the Division of Infectious Diseases, Massachusetts General Hospital, Cambridge, Massachusetts 02139.
authors.forename: Leigh A
authors.initials: LA
authors.lastname: Baxt
authors.affiliation: From the Division of Infectious Diseases, Massachusetts General Hospital, Cambridge, Massachusetts 02139.
authors.forename: Bobby Brooke
authors.initials: BB
authors.lastname: Herrera
authors.affiliation: From the Division of Infectious Diseases, Massachusetts General Hospital, Cambridge, Massachusetts 02139.
authors.forename: Emily
authors.initials: E
authors.lastname: Kahoud
authors.affiliation: Systems Biology, Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: Gavin
authors.initials: G
authors.lastname: MacBeath
authors.affiliation: From the Division of Infectious Diseases, Massachusetts General Hospital, Cambridge, Massachusetts 02139, Departments of Microbiology and Immunobiology and marcia.goldberg@mgh.harvard.edu.
authors.forename: Marcia B
authors.initials: MB
authors.lastname: Goldberg
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry* / Q000378:metabolism*; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D017124:Conserved Sequence; D003598:Cytoskeletal Proteins / Q000737:chemistry* / Q000378:metabolism*; D022001:Focal Adhesions / Q000378:metabolism; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D042541:Intracellular Space / Q000382:microbiology; D060588:LIM Domain Proteins / Q000737:chemistry* / Q000378:metabolism*; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D040081:Protein Array Analysis; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D011493:Protein Kinase C / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D012760:Shigella; D015398:Signal Transduction
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D001426:Bacterial Proteins; D003598:Cytoskeletal Proteins; D060588:LIM Domain Proteins; D050505:Mutant Proteins; C484380:PDLIM7 protein, human; D010455:Peptides; D011493:Protein Kinase C
keywords: Bacterial Pathogenesis; OspE; PDLIM Proteins; PDZ Domain; Protein Kinase C (PKC); Protein-Protein Interaction; Secretion; Shigella; Type III Secretion System (T3SS)