title: Human heat shock protein (Hsp) 90 interferes with Neisseria meningitidis adhesin A (NadA)-mediated adhesion and invasion.
abstract: NadA (N eisseria meningitidisadhesin A), a meningococcal surface protein, mediates adhesion to and invasion of human cells, an activity in which host membrane proteins have been implicated. While investigating these host factors in human epithelial cells by affinity chromatography, we discovered an unanticipated interaction of NadA with heat shock protein (Hsp) 90, a molecular chaperone. The specific in vitro interaction of recombinant soluble NadA and Hsp90 was confirmed by co-immunoprecipitations, dot and far-Western blot. Intriguingly, ADP, but not ATP, was required for this association, and the Hsp90 inhibitor 17-AAG promoted complex formation. Hsp90 binding to an Escherichia coli strain used as carrier to express surface exposed NadA confirmed these results in live bacteria. We also examined RNA interference, plasmid-driven overexpression, addition of exogenous rHsp90 and 17-AAG inhibition in human epithelial cells to further elucidate the involvement of Hsp90 in NadA-mediated adhesion and invasion. Together, these data suggest an inverse correlation between the amount of host Hsp90 and the NadA adhesive/invasive phenotype. Confocal microscopy also demonstrated that meningococci interact with cellular Hsp90, a completely novel finding. Altogether our results show that variation of host Hsp90 expression or activity interferes with adhesive and invasive events driven by NadA.
authors: Research Center, Novartis Vaccines and Diagnostics, Siena, Italy.
authors: Paolo
authors: P
authors: Montanari
authors: Giuseppe
authors: G
authors: Bozza
authors: Barbara
authors: B
authors: Capecchi
authors: Elena
authors: E
authors: Caproni
authors: Riccardo
authors: R
authors: Barrile
authors: Nathalie
authors: N
authors: Norais
authors: Mirco
authors: M
authors: Capitani
authors: Michele
authors: M
authors: Sallese
authors: Paola
authors: P
authors: Cecchini
authors: Laura
authors: L
authors: Ciucchi
authors: Zhenai
authors: Z
authors: Gao
authors: Rino
authors: R
authors: Rappuoli
authors: Mariagrazia
authors: M
authors: Pizza
authors: Beatrice
authors: B
authors: Aricò
authors: Marcello
authors: M
authors: Merola
mesh_terms: D018829:Adhesins, Bacterial / Q000378:metabolism*; D000595:Amino Acid Sequence; D001422:Bacterial Adhesion; D016227:Benzoquinones / Q000494:pharmacology; D002478:Cells, Cultured; D004926:Escherichia coli / Q000235:genetics / Q000502:physiology; D018841:HSP90 Heat-Shock Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D054884:Host-Pathogen Interactions; D006801:Humans; D047029:Lactams, Macrocyclic / Q000494:pharmacology; D008589:Meningococcal Infections / Q000378:metabolism / Q000382:microbiology; D008969:Molecular Sequence Data; D009345:Neisseria meningitidis / Q000502:physiology*; D011485:Protein Binding; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D018829:Adhesins, Bacterial; D016227:Benzoquinones; D018841:HSP90 Heat-Shock Proteins; D047029:Lactams, Macrocyclic; D011994:Recombinant Proteins; C112765:tanespimycin