title: PscI is a type III secretion needle anchoring protein with in vitro polymerization capacities.
abstract: The export of bacterial toxins across the bacterial envelope requires the assembly of complex, membrane-embedded protein architectures. Pseudomonas aeruginosa employs type III secretion (T3S) injectisome to translocate exotoxins directly into the cytoplasm of a target eukaryotic cell. This multi-protein channel crosses two bacterial membranes and extends further as a needle through which the proteins travel. We show in this work that PscI, proposed to form the T3S system (T3SS) inner rod, possesses intrinsic properties to polymerize into flexible and regularly twisted fibrils and activates IL-1β production in mouse bone marrow macrophages in vitro. We also found that point mutations within C-terminal amphipathic helix of PscI alter needle assembly in vitro and T3SS function in cell infection assays, suggesting that this region is essential for an efficient needle assembly. The overexpression of PscF partially compensates for the absence of the inner rod in PscI-deficient mutant by forming a secretion-proficient injectisome. All together, we propose that the polymerized PscI in P. aeruginosa optimizes the injectisome function by anchoring the needle within the envelope-embedded complex of the T3S secretome and - contrary to its counterpart in Salmonella - is not involved in substrate switching.
authors.affiliation: INSERM, UMR-S 1036, Biology of Cancer and Infection, Grenoble, France; CNRS, Bacterial Pathogenesis and Cellular Responses, ERL 5261, Grenoble, France; Université Grenoble Alpes, F-38041, Grenoble, France; CEA, DSV/iRTSV, F-38054, Grenoble, France.
authors.forename: Laura
authors.initials: L
authors.lastname: Monlezun
authors.forename: David
authors.initials: D
authors.lastname: Liebl
authors.forename: Daphna
authors.initials: D
authors.lastname: Fenel
authors.forename: Teddy
authors.initials: T
authors.lastname: Grandjean
authors.forename: Alice
authors.initials: A
authors.lastname: Berry
authors.forename: Guy
authors.initials: G
authors.lastname: Schoehn
authors.forename: Rodrigue
authors.initials: R
authors.lastname: Dessein
authors.forename: Eric
authors.initials: E
authors.lastname: Faudry
authors.forename: Ina
authors.initials: I
authors.lastname: Attree
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D053583:Interleukin-1beta / Q000235:genetics / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism / Q000382:microbiology; D051379:Mice; D058105:Polymerization; D021381:Protein Transport; D011552:Pseudomonas Infections / Q000235:genetics / Q000378:metabolism / Q000382:microbiology*; D011550:Pseudomonas aeruginosa / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000069296:Type III Secretion Systems / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D053583:Interleukin-1beta; D000069296:Type III Secretion Systems