Project description:Microarray comparative genome hybridization (mCGH) data was collected from one Neisseria cinerea, two Neisseria lactamica, two Neisseria gonorrhoeae, and 48 Neisseria meningitidis isolates. For N. meningitidis, these isolates are from diverse clonal complexes, invasive and carriage strains, and all major serogroups. The microarray platform represented N. meningitidis strains MC58, Z2491, and FAM18 and N. gonorrhoeae FA1090.
Project description:Neisseria gonorrhoeae (NG) exhibits high genome plasticity caused by an unusually high density and diversity of transposable elements, and easily performs various mechanisms of drug resistance. Here we investigated the i19.05 clinical isolate with reduced susceptibility to penicillin (MIC=0.5 mg/L), tetracycline (MIC=0.5 mg/L), and azithromycin (MIC=1.0 mg/L), which carried no known genetic resistance determinants except of penA, which cannot explain the expression of the resistant phenotype. In addition, it attracted our attention to the presence of a new and unique mutation of Asn105Ser in SurA and several mutations in Omp85 (BamA). The goal of our study was to search for new molecular mechanisms of drug resistance. The pan susceptible n01.08 NG clinical isolate was involved as a control to compare, as well as a recipient in transformation procedure. The fragments of i19.05 genome contained mutant surA, omp85, and penA genes were amplified and used in spot-transformation of the n01.08 recipient isolate as described (Ilina, 2013). Finally, a resistant transformant NG05 (PenAmut, Ompmut, SurAmut) was obtained. For comprehensive proteomic analysis via LC-MS/MS, the proteins from the all tested N. gonorrhoeae strains were fractionated on cell envelope (CE) (including outer membrane, periplasmic, inner membrane) and cytosol (C). A total of 1125 proteins in the CE fraction, of which 894 were common in all strains were identified. Proteomics of the C fraction in the same experiment yielded a total of 928 proteins, of which 676 were shared among all strains. Proteome coverage for both fractions ranged from 52.72% (1111 proteins) in n01.08 to 54.53% (1149 proteins) in i19.05.
Project description:The sexually transmitted pathogen Neisseria gonorrhoeae releases outer membrane vesicles (OMVs) during infections. OMVs traffic the major porin PorB, other membrane proteins and lipo-oligosaccharide (LOS) into host innate immune cells and activate programmed cell death pathways and inflammation. Little is known, however, about the proteome and LOS content of OMVs released by clinical strains isolated from different infection sites, and whether this affects immune responses. Here, we characterized OMVs from four N. gonorrhoeae isolates and determined their size, abundance, proteome and activation of inflammatory responses in macrophages. The overall proteome of the OMVs was conserved between the four different isolates, included major outer membrane, periplasm, cytoplasmic membrane proteins. Despite this, we observed differences in the rate of OMV biogenesis and the relative abundance of major outer membrane proteins and LOS. Consequently, OMVs from clinical isolates induced varying rates of macrophage cell death and the secretion of interleukin-1 family members, such as Il-1andIl-1. Overall, these findings demonstrate that clinical isolates of N. gonorrhoeae utilize OMVs to release major proteins and lipids, which affects innate immune responses.
2024-08-27 | PXD042787 | Pride
Project description:Genomic epidemiology and antimicrobial resistance in Neisseria gonorrhoeae isolates from Ghana
Project description:Neisseria gonorrhoeae is the causative agent of gonorrhea, a leading sexually transmitted disease with severe complications on reproductive health. The U.S. Centers for Disease Control and Prevention has categorized the public health threat induced by N. gonorrhoeae as “urgent”, due to the ease of transmission and the fast emergence of multi-drug resistant strains. The need for development of vaccines and understanding the underlying factors leading to antibiotic resistance is of utmost importance. The proteomic profiles of the 14 WHO N. gonorrhoeae reference strains have been compared to the WHO F reference strain using a mass spectrometry with tandem mass tags (TMT) labeling to analyze the cell envelope and the cytoplasmic fractions extracted from each strain. Identifying novel vaccine candidates and proteomic signatures for antimicrobial resistance will further our understanding of N. gonorrhoeae proteotypes, in relationship to their respective genotypes and phenotypes, and provide deep insights that will impact the development of preventive and therapeutic tools to combat gonorrhea.
Project description:Comparison of transcriptional profiling between the 3 Neisseria meningitidis strains [serogroup A (Z2491), Serogroup B (MC58), and Serogroup C (FAM18)] and the 2 Neisseria gonorrhoeae strain (FA1090 and MS11).