Contribution of the gonococcal NEIS1446-ispD gene conversion to the pathobiology of the Neisseria meningitidis urethritis clade, NmUC.

Over the last decade, a Neisseria meningitidis (Nm) urethritis-causing clade (NmUC) has emerged to cause clusters of meningococcal urethritis in the United States and globally. One genomic signature of NmUC is the integration of Neisseria gonorrhoeae (Ng) DNA in an operon, NEIS1446-NEIS1438, which partially replaced the Nm ispD gene. IspD is the 2-C-methyl-d-erythritol 4-phosphate cytidylyltransferase of the terpenoid precursor synthesis pathway, required for the production of ubiquinones of the electron transfer chain. IspD is essential in several gram-negative bacteria. The biological importance of the NEIS1446-ispD gene conversion event for NmUC was investigated. The ispD gene was found to be essential in NmUC (CNM3) and non-clade Nm (MC58), and a mutation at the native locus can only be made with the insertion of a second ispD copy in the genome. The IspD_MC58 variant was more efficient at promoting aerobic growth at a low level than IspD_CNM3; the two proteins differ by 15 residues. Maximal aerobic growth densities of strains with an NmUC background resembled Ng (FA19), and both were significantly lower than Nm. In contrast to non-clade Nm, all NmUC strains survived well anaerobically. Increasing ispD expression by titrating IPTG in non-clade Nm enhanced anaerobic survival. Translational reporters of the NmUC and Ng promoters demonstrated similar expression levels, and both were significantly higher than non-clade Nm, under aerobic and microaerobic conditions. Our findings suggest that the integration of gonococcal DNA into the NEIS1446-NEIS1438 operon of NmUC has increased ispD expression, contributing to NmUC's adaptation to the oxygen-limited environment of the human urogenital tract.