Construction of a complete set of Neisseria meningitidis mutants and its use for the phenotypic profiling of this human pathogen

Abstract

The bacterium Neisseria meningitidis causes life-threatening meningitis and sepsis. Here, we construct a complete collection of defined mutants in protein-coding genes of this organism, identifying all genes that are essential under laboratory conditions. The collection, named NeMeSys 2.0, consists of individual mutants in 1584 non-essential genes. We identify 391 essential genes, which are associated with basic functions such as expression and preservation of genome information, cell membrane structure and function, and metabolism. We use this collection to shed light on the functions of diverse genes, including a gene encoding a member of a previously unrecognised class of histidinol-phosphatases; a set of 20 genes required for type IV pili function; and several conditionally essential genes encoding antitoxins and/or immunity proteins. We expect that NeMeSys 2.0 will facilitate the phenotypic profiling of a major human bacterial pathogen.

Neisseria genes required for persistence identified via in vivo screening of a transposon mutant library

“…The curation yielded a final set of 776 putative housekeeping genes with a lower than anticipated gene insertion index and 192 genes without detectable insertions in library passage and inoculum samples (S1 Data). Annotation, pathway mapping and over representation analysis revealed their functions were predominately associated with central and secondary metabolism, ribosomal structure and biogenesis, cell envelope biosynthesis, cell division, and nutrient acquisition and transport (S1B and S1C Fig) . The prevalence of genes associated with vital physiologic processes, including those known to be essential in other organisms, (e.g., signal peptidase 1, ribosomal proteins, NrdB, elongation factor Tu) supports our notion that insertions in these open reading frames convey a defect in in vitro growth [44].…”

“…gluP encodes a glucose/galactose MFS family transporter with similarity to Brucella abortus GluP PLOS PATHOGENS (54.5% identity over 400 amino acids). While gluP has been identified as differentially expressed in N. meningitidis in vitro infection studies, its exact contribution to persistent carriage remains unknown in this context [44,82]. B. abortus gluP is essential for glucose uptake in host environments, as mutants display reduced survival in alternatively activated macrophages and cannot persist in a chronic mouse infection model [83,84].…”

Mechanism of assembly of type 4 filaments: everything you always wanted to know (but were afraid to ask)

“…T4aP are the best characterized T4F. The two major diderm models are P. aeruginosa and N. meningitidis, in which exhaustive genetic screens have identified all the genes involved in T4aP biology [64,65]. The genes necessary for pilus biogenesis -denoted by the same pil mnemonic in these two species, but often with different letters -are scattered through the genome.…”

Variation in supplemental carbon dioxide requirements defines lineage-specific antibiotic resistance acquisition in Neisseria gonorrhoeae

“…Carbonic anhydrases have been previously shown to be targets of sulfa antibiotics 12 and to mediate natural competence in N. meningitidis 21 . Sulfa antibiotics were one of the first antibiotics to be used to treat gonorrhea 22 and primarily target dihydropteroate synthase in the folate synthesis pathway.…”