[show abstract][hide abstract] ABSTRACT: Phase variable expression, mediated by high frequency reversible changes in the length of simple sequence repeats, facilitates adaptation of bacterial populations to changing environments and is frequently important in bacterial virulence. Here we elucidate a novel phase variable mechanism for NadA, an adhesin and invasin of Neisseria meningitidis. The NadR repressor protein binds to operators flanking the phase variable tract and contributes to the differential expression levels of phase variant promoters with different numbers of repeats likely due to different spacing between operators. We show that IHF binds between these operators, and may permit looping of the promoter, allowing interaction of NadR at operators located distally or overlapping the promoter. The 4-hydroxyphenylacetic acid, a metabolite of aromatic amino acid catabolism that is secreted in saliva, induces NadA expression by inhibiting the DNA binding activity of the repressor. When induced, only minor differences are evident between NadR-independent transcription levels of promoter phase variants and are likely due to differential RNA polymerase contacts leading to altered promoter activity. Our results suggest that NadA expression is under both stochastic and tight environmental-sensing regulatory control, both mediated by the NadR repressor, and may be induced during colonization of the oropharynx where it plays a major role in the successful adhesion and invasion of the mucosa. Hence, simple sequence repeats in promoter regions may be a strategy used by host-adapted bacterial pathogens to randomly switch between expression states that may nonetheless still be induced by appropriate niche-specific signals.
[show abstract][hide abstract] ABSTRACT: Neisseria gonorrhoeae displays considerable potential for antigenic variation as shown in human experimental studies. Various surface antigens can change either by antigenic variation using RecA-dependent recombination schemes (e.g. PilE antigenic variation) or, alternatively, through phase variation (on/off switching) in a RecA-independent fashion (e.g. Opa and lipooligosaccharide phase variation). PilE antigenic variation has been well documented over the years. However, with the availability of the N. gonorrhoeae FA1090 genome sequence, considerable genetic advances have recently been made regarding the mechanistic considerations of the gene conversion event, leading to an altered PilE protein. This review will compare the various models that have been presented and will highlight potential mechanistic problems that may constrain any genetic model for pilE gene variation.
[show abstract][hide abstract] ABSTRACT: The role of the RecBCD recombination pathway in PilE antigenic variation in Neisseria gonorrhoeae is contentious and appears to be strain dependent. In this study, N. gonorrhoeae strain MS11 recB mutants were assessed for recombination/repair. MS11 recB mutants were found to be highly susceptible to DNA treatments that caused double-chain breaks and were severely impaired for growth; recB growth suppressor mutants arose at high frequencies. When the recombination/repair capacity of strain MS11 was compared to that of strains FA1090 and P9, innate differences were observed between the strains, with FA1090 and P9 rec(+) bacteria presenting pronounced recombination/repair defects. Consequently, MS11 recB mutants present a more robust phenotype than the other strains that were tested. In addition, MS11 recB mutants are also shown to be defective for pilE/pilS recombination. Moreover, pilE/pilS recombination is shown to proceed with gonococci that carry inverted pilE loci. Consequently, a novel RecBCD-mediated double-chain-break repair model for PilE antigenic variation is proposed.
Journal of bacteriology 12/2007; 189(22):7983-90. · 3.94 Impact Factor
[show abstract][hide abstract] ABSTRACT: The bacterial stringent response is a pleiotrophic physiological response that is evoked when bacteria are subjected to nutrient stress and is mediated through the accumulation of hyperphosphorylated guanine nucleotides ((p)ppGpp) which are synthesized by the combined action of the relA and spoT gene products. The relA and spoT genes were cloned from Neisseria gonorrhoeae strain MS11 and various insertional and deletion mutants were constructed. Deletion of the gonococcal relA gene abrogated the production of (p)ppGpp when the organism was starved for the amino acid serine. Also, N. gonorrhoeaeDeltarelA null mutants were impaired for growth when propagated on rich medium, a phenotype that could be relieved by deleting the spoT gene. Sequence analysis of the gonococcal SpoT polypeptide indicated a strong similarity to its Escherichia coli counterpart. However, in contrast to studies with E. coli, insertional spoT mutants could be obtained that still accumulated (p)ppGpp when gonococci were starved for nutrients provided that the non-polar insertions were located downstream of the putative phosphohydrolase active site. In time course studies, it is also shown that gonococci rapidly accumulate (p)ppGpp (within 5 min) when encountering nutrient deprivation.