title: Construction and characterization of a derivative of Neisseria gonorrhoeae strain MS11 devoid of all opa genes.
abstract: To better understand the role of Opa in gonococcal infections, we created and characterized a derivative of MS11 (MS11Δopa) that had the coding sequence for all 11 Opa proteins deleted. The MS11Δopa bacterium lost the ability to bind to purified lipooligosaccharide (LOS). While nonpiliated MS11Δopa and nonpiliated Opa-expressing MS11 cells grew at the same rate, nonpiliated MS11Δopa cells rarely formed clumps of more than four bacteria when grown in broth with vigorous shaking. Using flow cytometry analysis, we demonstrated that MS11Δopa produced a homogeneous population of bacteria that failed to bind monoclonal antibody (MAb) 4B12, a MAb specific for Opa. Opa-expressing MS11 cells consisted of two predominant populations, where ∼85% bound MAb 4B12 to a significant level and the other population bound little if any MAb. Approximately 90% of bacteria isolated from a phenotypically Opa-negative colony (a colony that does not refract light) failed to bind MAb 4B12; the remaining 10% bound MAb to various degrees. Piliated MS11Δopa cells formed dispersed microcolonies on ME180 cells which were visually distinct from those of piliated Opa-expressing MS11 cells. When Opa expression was reintroduced into MS11Δopa, the adherence ability of the strain recovered to wild-type levels. These data indicate that Opa contributes to both bacterium-bacterium and bacterium-host cell interactions.
authors.affiliation: University of Maryland, Department of Cell Biology and Molecular Genetics, College Park, Maryland, USA.
authors.forename: Adriana
authors.initials: A
authors.lastname: LeVan
authors.forename: Lindsey I
authors.initials: LI
authors.lastname: Zimmerman
authors.forename: Amanda C
authors.initials: AC
authors.lastname: Mahle
authors.forename: Karen V
authors.initials: KV
authors.lastname: Swanson
authors.forename: Philip
authors.initials: P
authors.lastname: DeShong
authors.forename: Juhee
authors.initials: J
authors.lastname: Park
authors.forename: Vonetta L
authors.initials: VL
authors.lastname: Edwards
authors.forename: Wenxia
authors.initials: W
authors.lastname: Song
authors.forename: Daniel C
authors.initials: DC
authors.lastname: Stein
mesh_terms: D000907:Antibodies, Bacterial / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology; D001422:Bacterial Adhesion; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D017353:Gene Deletion; D005816:Genetic Complementation Test; D008070:Lipopolysaccharides / Q000378:metabolism; D009344:Neisseria gonorrhoeae / Q000235:genetics* / Q000254:growth & development / Q000502:physiology; D011485:Protein Binding
chemical_list: D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; D008070:Lipopolysaccharides; C023023:lipid-linked oligosaccharides; C090348:Opa protein, Neisseria