title: Loss of co-chaperone TopJ impacts adhesin P1 presentation and terminal organelle maturation in Mycoplasma pneumoniae.
abstract: Mycoplasma pneumoniae is a wall-less human respiratory tract pathogen that colonizes mucosal epithelium via a polar terminal organelle having a central electron-dense core and adhesin-related proteins clustered at a terminal button. A mutant lacking J-domain co-chaperone TopJ is non-cytadherent and non-motile, despite having a core and normal levels of the major cytadherence-associated proteins. J-domain co-chaperones work with DnaK to catalyse polypeptide binding and subsequent protein folding. Here we compared features of the topJ mutant with other cytadherence mutants to elucidate the contribution of TopJ to cytadherence function. The topJ mutant was similar ultrastructurally to a non-cytadherent mutant lacking terminal organelle proteins B/C, including aberrant core positioning and cell morphology in thin sections, but exhibited a hybrid satellite growth pattern with features of mutants both having and lacking a core. Time-lapse images of mycoplasmas expressing a YFP fusion with terminal organelle protein P41 suggested that terminal organelle formation/positioning was delayed or poorly co-ordinated with cell growth in the absence of TopJ. TopJ required a core for localization, perhaps involving HMW1. P1 trypsin accessibility on other non-cytadherent mutants was significantly enhanced over wild type but unexpectedly was reduced with topJ mutant cells, suggesting impaired processing, translocation and/or folding of this adhesin.
authors.affiliation: Department of Microbiology, University of Georgia, Athens, GA 30602, USA.
authors.forename: Jason M
authors.initials: JM
authors.lastname: Cloward
authors.forename: Duncan C
authors.initials: DC
authors.lastname: Krause
mesh_terms: D018829:Adhesins, Bacterial / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D055786:Gene Knockout Techniques; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism*; D009177:Mycoplasma pneumoniae / Q000254:growth & development* / Q000378:metabolism* / Q000648:ultrastructure; D015388:Organelles / Q000378:metabolism* / Q000648:ultrastructure; D059008:Time-Lapse Imaging
chemical_list: D018829:Adhesins, Bacterial; D001426:Bacterial Proteins; D018832:Molecular Chaperones; C035471:adhesin, Mycoplasma pneumoniae