Project description:We calculated half-life values of mRNAs quantified by RNA-Seq by a suitable method of normalization. We determined the half-lives of more than 2200 mRNAs in the Stenotrophomonas maltophilia D457 wild-type strain and in an isogenic RNase G deficient mutant. Median half-lives were 2,74 and 3 min in the wild-type and the rng-deficient mutant respectively. We found an overall enhancement of half-life times of mRNAs when the gene encoding RNase G is lacking, showing that many RNAs are targets of RNase G in S. maltophilia. For achieving such goal, we propose a method for the normalization of RNA-Seq based studies on global bacterial mRNA decay.
Project description:Transcriptomic analysis of S. maltophilia D457 after one-hour induction with the antibiotic fosfomycin, the intermediate metabolites phosphoenolpyruvate or glyceraldehyde-3-phosphate
Project description:Stenotrophomonas maltophilia is an important opportunistic pathogen affecting primarily hospitalized and immuno-compromised hosts. We constructed an hfq deletion mutant (Delta-hfq) of S. maltophilia, and compared the behaviour of wild-type and Delta-hfq S. maltophilia cells in a variety of assays. Differential RNA sequencing analysis (dRNA-seq) of RNA isolated from S. maltophilia wild-type and Delta-hfq strains showed that Hfq regulates expression of genes encoding flagellar and fimbrial components, transmembrane proteins, as well as enzymes involved in different metabolic pathways. Moreover, we analysed expression of several sRNAs identified by dRNA-seq in wild-type. The accumulation of two sRNAs was strongly reduced in the absence of Hfq. TEX (terminator exonuclease) treated and untreated libraries of the wild type and the Delta-hfq mutant were sequenced and compared
Project description:We sought to determine how a cystic fibrosis isolate of Stenotrophomonas maltophilia responds to relevant pH gradients (pH 5, 7, and 9) by growing the bacterium in phosphate buffered media and conducting RNAseq experiments. Our data suggests acidic conditions are stressful for strain FLR19, as it responded by increasing expression of stress-response and antibiotic-resistance genes.
Project description:Stenotrophomonas maltophilia is an important opportunistic pathogen affecting primarily hospitalized and immuno-compromised hosts. We constructed an hfq deletion mutant (Delta-hfq) of S. maltophilia, and compared the behaviour of wild-type and Delta-hfq S. maltophilia cells in a variety of assays. Differential RNA sequencing analysis (dRNA-seq) of RNA isolated from S. maltophilia wild-type and Delta-hfq strains showed that Hfq regulates expression of genes encoding flagellar and fimbrial components, transmembrane proteins, as well as enzymes involved in different metabolic pathways. Moreover, we analysed expression of several sRNAs identified by dRNA-seq in wild-type. The accumulation of two sRNAs was strongly reduced in the absence of Hfq.
Project description:The goal of this study was to elucidate genes that are employed by the bacterivorous nematode Caenorhabditis elegans to respond to the emerging nosocomial bacterial pathogen Stenotrophomonas maltophilia.
Project description:Stenotrophomonas maltophilia is an emerging multidrug resistance opportunistic pathogen affecting immunocompromised and hospitalized patients. S. maltophilia is an environmental bacterium which adapts to human body and causing infection. S. rhizophilia, a non-pathogenic and phylogenetic neighbour of S. maltophilia is unable to grow at human body temperature. Thus, to understand molecular mechanism underlying the adaptation of S. maltophilia at human body temperature, we performed the comparative transcriptome analysis of S.maltophilia at 28 °C (representative for the environmental niches) and 37 °C (representative for human body) by using RNA-Seq. The major temperature-induced genes include genes for Type IV secretion system, aerotaxis, and cation diffusion facilitator family transporter suggesting its potential role in the adaptation and virulence of S. maltophilia. The downregulated genes at 37 °C includes the genes for the cell motility, energy generation and metabolism, lipid metabolism, translation, amino acid metabolism and transport, replication and repair, inorganic ion and transport metabolism lipid metabolism, coenzyme metabolism.