title: Type III secretion chaperones of Pseudomonas syringae protect effectors from Lon-associated degradation.
abstract: The hrp type III secretion system (TTSS) of Pseudomonas syringae translocates effector proteins into the cytoplasm of host cells. Proteolysis of HrpR by Lon has been shown to negatively regulate the hrp TTSS. The inability to bypass Lon-associated effects on the regulatory system by ectopic expression of the known regulators suggested a second site of action for Lon in TTSS-dependent effector secretion. In this study we report that TTSS-dependent effectors are subject to the proteolytic degradation that appears to be rate-limiting to secretion. The half-lives of the effectors AvrPto, AvrRpt2, HopPsyA, HopPsyB1, HopPtoB2, HopPsyV1, HopPtoG and HopPtoM were substantially higher in bacteria lacking Lon. TTSS-dependent secretion of several effectors was enhanced from Lon mutants. A primary role for chaperones appears to be protection of effectors from Lon-associated degradation prior to secretion. When coexpressed with their cognate chaperone, HopPsyB1, HopPsyV1 and HopPtoM were at least 10 times more stable in strains expressing Lon. Distinct Lon-targeting and chaperone-binding domains were identified in HopPtoM. The results imply that Lon is involved at two distinct levels in the regulation of the P. syringae TTSS: regulation of assembly of the secreton and modulation of effector secretion.
authors.affiliation: Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD 20742, USA.
authors.forename: Liliana C
authors.initials: LC
authors.lastname: Losada
authors.forename: Steven W
authors.initials: SW
authors.lastname: Hutcheson
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D018832:Molecular Chaperones / Q000378:metabolism*; D010935:Plant Diseases / Q000382:microbiology; D018515:Plant Leaves / Q000382:microbiology; D049070:Protease La / Q000378:metabolism*; D044224:Pseudomonas syringae / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity*; D012808:Sigma Factor / Q000378:metabolism; D014026:Nicotiana / Q000382:microbiology
chemical_list: D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; C085855:HrpL protein, Pseudomonas syringae; D018832:Molecular Chaperones; D012808:Sigma Factor; D049070:Protease La