Mutagenesis of the Neisseria gonorrhoeae porin reduces invasion in epithelial cells and enhances phagocyte responsiveness.
Porin (PorB), the major outer membrane protein of Neisseria gonorrhoeae, has been implicated in pathogenesis previously. However, the fact that porin deletion mutants are not viable has complicated investigations. Here, we describe a method of manipulating the porin gene site-specifically. N. gonorrhoeae MS11, which harbours the porB1B (P.1B) porin allele, was used to generate mutants carrying deletions in the surface loops 1 and 5. An 11-amino-acid deletion in loop 1 impaired Opa50-dependent invasion into human Chang epithelial cells, whereas loop 5 deletion exhibited no apparent phenotype. In a second approach, the complete gonococcal porB1B was replaced by the porBNia gene of Neisseria lactamica. Such mutants were unable to induce efficient uptake by epithelial cells but induced an enhanced respiratory response in HL60 phagocytic cells. The increased respiratory burst was accompanied by an enhanced phagocytic uptake of the mutant compared with the wild-type strain. Our data extend previous evidence for multiple central functions of PorB in the infection process.
Max-Planck-Institut für Biologie, Abt Infektionsbiologie, Tübingen, Germany.
F J
FJ
Bauer
T
T
Rudel
M
M
Stein
T F
TF
Meyer
D000483:Alleles; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D002448:Cell Adhesion; D004847:Epithelial Cells / Q000382:microbiology*; D005822:Genetic Vectors; D005838:Genotype; D018922:HL-60 Cells / Q000378:metabolism; D006801:Humans; D008163:Luminescent Measurements; D016296:Mutagenesis; D009344:Neisseria gonorrhoeae / Q000235:genetics*; D009841:Oligonucleotides; D010586:Phagocytes / Q000502:physiology; D018272:Porins / Q000235:genetics* / Q000502:physiology; D016897:Respiratory Burst / Q000502:physiology; D013997:Time Factors
D001425:Bacterial Outer Membrane Proteins; D009841:Oligonucleotides; D018272:Porins; C057699:porin protein, Neisseria