Project description:Directional RNAseq analyses was undertaken in Klebsiella pneumoniae Ecl8 and isogenic mutants Ecl8delta ramA and Ecl8delta ramR to determine the RamA regulon. All samples were grown in Luria Bertani broth until OD600 approx 0.6 prior to RNA extraction. Differential expression was determined using DEseq upon pairwise comparisons of Ecl8 vs Ecl8delta ramA, Ecl8deltaramA vs Ecl8deltaramR, Ecl8 vs Ecl8deltaramR.This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/
Project description:Purpose: we conducted this study to investigate whether gut K.pn is a causal factor for HTN pathogenesis and the potential mechanism Methods: Renal transcriptome changes were assessed by RNA-sequencing Results: significant upregulation of several genes related to HTN pathogenesis were observed Conclusions:These results provide evidence that the enrichment of K.pn acts as a direct contributor for hypertension pathogenesis
Project description:Klebsiella pneumoniae poses significant threat to global health. Detailed investigation to the nature of its interaction to human epithelial cells is still lacking. OmpR is an important regulator to the expression of major outer membrane protein genes, ompF and ompC. Here, we exploited the recently described dual RNA-seq to simultaneously measure genome-wide expression of host and pathogen two hours into infection. By comparing OmpR deleted strain to its wildtype parental strain, we, simultaneously, reconstructed OmpR regulon and deletion effect to host response.
Project description:In bacteria the defence system to counter oxidative stress is orchestrated by three transcriptional factors – SoxS, SoxR and OxyR. Although the transcriptional regulon of these factors are known in many bacteria, similar data is not available for K. pneumoniae. To address this data gap, oxidative stress was induced in K. pneumoniae MGH 78578 using paraquat and the corresponding regulon was identified using RNA-seq. Since soxS was significantly induced , a soxS mutant was constructed to decipher this regulon in K. pneumoniae MGH75878. The ‘oxidative SoxS regulon’, comprising common genes differentially regulated genes in oxidative and soxS regulon was identified from both regulons – characterised a stringent group of genes which were regulated by SoxS during oxidative stress. Efflux pump encoding genes like acrAB-tolC, acrE along with marRABwere identified in the oxidative SoxS regulon. The phenotypic effect of the observed efflux pump regulation was confirmed in the soxS mutant that exhibited an 2 fold reduction in the minimum bactericidal concentration (MBC) against tetracycline compared to that of the isogenic wild type. Impaired efflux activity, allowing tetracycline to be accumulated in the cytoplasm to bactericidal levels, was further confirmed using tetraphenylphosphonium (TPP+) ion accumulation assay. The susceptibility of the soxS mutant against tetracycline was also apparent in vivo, in the zebrafish embryo model. We conclude that the soxS gene could be considered as a genetic target against which an inhibitor could be developed and be used in combinatorial therapy with tetracycline to combat infections associated with multi-drug resistant K. pneumoniae.
Project description:We conducted this study to investigate whether gut K.pn is a causal factor for HTN pathogenesis and the potential mechanism Ttranscriptome changes of aorta were assessed by RNA-sequencing
