Neisseria gonorrhoeae PLD directly interacts with Akt kinase upon infection of primary, human, cervical epithelial cells.
Neisseria gonorrhoeae secrets a phospholipase D (NgPLD), which augments complement receptor 3 (CR3)-mediated invasion of cervical epithelial cells. To elucidate the signalling pathways triggered with gonococcus CR3-engagement and the putative function of NgPLD in these events, we analysed the contribution of the phosphoinositide-Akt pathway to cervical infection. Our data indicated that Akt plays a critical role in cervical infection. Inhibition of myosin light chain kinase, PtdIns(4,5)P2, and Akt functions resulted in decreased gonococcus invasion of primary, human, cervical epithelial cells as well as Akt kinase activity. Akt activity was similarly impaired when cervical cells were challenged with NgPLD-mutant gonococci. Conversely, the PI3-kinase inhibitor, LY294002, enhanced gonococcal invasion of, and Akt activity within, primary cervical cells. We demonstrated that NgPLD directly binds to the Akt PH domain and can compete with a natural Akt ligand, PtdIns(3,4,5)P3, for Akt binding. Collectively, our data suggested that NgPLD augments gonococcus invasion of cervical epithelia by interacting with Akt kinase in a PI3-kinase-independent manner, which results in subversion of normal cervical cell signalling.
Edwards
Jennifer L
JL
Department of Microbiology, The University of Iowa, Iowa City, IA 52242, USA. edwardsj@ccri.net
Apicella
Michael A
MA
D001665:Binding Sites; D001667:Binding, Competitive; D002478:Cells, Cultured; D002584:Cervix Uteri / Q000201:enzymology* / Q000382:microbiology*; D004789:Enzyme Activation; D004847:Epithelial Cells / Q000201:enzymology / Q000382:microbiology; D005260:Female; D005798:Genes, Bacterial; D006069:Gonorrhea / Q000201:enzymology / Q000209:etiology / Q000382:microbiology; D006801:Humans; D066298:In Vitro Techniques; D009154:Mutation; D009344:Neisseria gonorrhoeae / Q000201:enzymology* / Q000235:genetics / Q000472:pathogenicity*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism; D010739:Phospholipase D / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D051057:Proto-Oncogene Proteins c-akt / Q000737:chemistry / Q000378:metabolism*; D015398:Signal Transduction; D014774:Virulence / Q000235:genetics / Q000502:physiology
D019869:Phosphatidylinositol 3-Kinases; D018129:Phosphatidylinositol Phosphates; D010739:Phospholipase D; D051057:Proto-Oncogene Proteins c-akt; C060974:phosphatidylinositol 3,4,5-triphosphate