Gonococcal invasion of epithelial cells driven by P.IA, a bacterial ion channel with GTP binding properties.
The neisserial porin P.I is a GTP binding protein that forms a voltage-gated channel that translocates into mammalian cell membranes and modulates host cell signaling events. Here, we report that P.I confers invasion of the bacterial pathogen Neisseria gonorrhoeae into Chang epithelial cells and that this event is controlled by GTP, as well as other phosphorus-containing compounds. Bacterial invasion was observed only for strains carrying the P.IA subtype of porin, which is typically associated with the development of disseminated neisserial disease, and did not require opacity outer membrane proteins, previously recognized as gonococcal invasins. Allelic replacement studies showed that bacterial invasiveness cotransferred with the P.IA (por1A) gene. Mutation of the P.I-associated protein Rmp did not alter the invasive properties. Cross-linking of labeled GTP to the porin revealed more efficient GTP binding to the P.IA than P.IB porin subtype. GTP binding was inhibited by an excess of unlabeled GTP, ATP, and GDP, as well as inorganic phosphate, but not by UTP or beta-glycerophosphate, fully in line with the respective invasion-inhibitory activities observed for these compounds. The P.IA-mediated cellular invasion may explain the more invasive behavior of P.IA strains in the natural infection and may broaden the basis for the development of a P.I-based gonococcal vaccine.
van Putten
J P
JP
Laboratory of Microbial Structure and Function, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, Hamilton, Montana 59840-2999, USA. jos_van_putten@nih.gov
Duensing
T D
TD
Carlson
J
J
D000483:Alleles; D000942:Antigens, Bacterial / Q000378:metabolism / Q000502:physiology*; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D002460:Cell Line; D004847:Epithelial Cells / Q000187:drug effects / Q000276:immunology* / Q000382:microbiology*; D019204:GTP-Binding Proteins / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D007473:Ion Channels / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D016254:Mutagenesis, Insertional; D009344:Neisseria gonorrhoeae / Q000187:drug effects / Q000472:pathogenicity*; D010710:Phosphates / Q000494:pharmacology; D018272:Porins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D007473:Ion Channels; D010710:Phosphates; D018272:Porins; C051591:Rmp protein, Neisseria; C050308:opacity proteins; C057699:porin protein, Neisseria; D019204:GTP-Binding Proteins