Project description:We reported the RNA-Seq results of Staphylococcus aureus Newman wild-type, walKD119A, walKV149A and DHBP-treated wild-type strains. We found that mutations of the potential signal transduction residues of WalK attenuate the activity of walKR two-component system, whereas DHBP supplementation activates this two-component system.
Project description:Genes that showed altered expression in Stk1 and Stp1 deficient Staphylococcus aureus Newman were identified. strain comparison, global regulation
Project description:To study the roles of NWMN_0641, we used microarray to compare the transcriptome of the NWMN_0641 deletion strain with that of the wild-type Staphylococcus aureus Newman strain. Transcriptome of the NWMN_0641 deletion mutant strain and the wild-type Newman strain
Project description:Array analysis of total RNA from broncho-alveolar lavage (BAL) from mice infected with influenza virus (PR8) and/or Staphylococcus aureus (Newman)
Project description:Staphylococcus aureus is a pathogen which can cause a wide range of infections. To find new targets for diagnostics and treatment as well as understanding host-pathogen interactions, many studies have focused on the bacterial surface proteins. In the study presented here, bacterial cell surface-shaving, using Lipid-based Protein Immobilization (LPI), followed by tandem mass tag (TMT) protocols for performing relative quantitative mass spectrometry proteomics, was performed to examine the surface proteome (surfaceome) of selected strains of S. aureus. The study included analyses of surface protein-deficient mutants compared to the wildtype S. aureus strain Newman, as well strain isolates of different clinical associations. Quantitative proteomics were applied in the analysis of the surface proteome of Staphylococcus aureus. In study 1, the differential abundance of proteins in the mutant strains ΔClfA, ΔSrtAΔSrtB and ΔSpa was compared to those in the Newman parental strain. All together 7880 peptides were identified in Study 1 corresponding to 1290 proteins, and the results clearly showed that the mutant strains were deficient in the knocked-out genes. The Clinical strains study (Study 2) included the Newman strain, the reference strains LS-1 and SH1000 and three clinical isolates, two from invasive infections and one from mild skin infection. A total of 4949 peptides were identified in Study 2 corresponding to 919 proteins. For each strain, approximately 20 proteins showed higher or lower abundance (fold changes) when compared to the Newman strain. The results indicate that surface shaving of intact bacteria by LPI in combination with protocols for performing quantitative proteomics makes it possible to distinguish differences in protein abundance of the surfaceome, including virulence factors, between S. aureus strains.
Project description:The SaeRS two-component regulatory system of Staphylococcus aureus is known to affect the expression of many genes. The SaeS protein is the histidine kinase responsible for phosphorylation of the response regulator SaeR. In S. aureus Newman, the sae system is constitutively expressed due to a point mutation in saeS, relative to other S. aureus strains, which results in substitution of proline for leucine at amino acid 18. Strain Newman is unable to form a robust biofilm and we report here that the biofilm-deficient phenotype is due to the saeSP allele. Replacement of the Newman saeSP with saeSL, or deletion of saeRS, resulted in a biofilm-proficient phenotype. Newman culture supernatants were observed to inhibit biofilm formation by other S. aureus strains, but did not affect biofilm formation by S. epidermidis. Culture supernatants of Newman saeSL or Newman ΔsaeRS had no significant effect on biofilm formation. The inhibitory factor was inactivated by incubation with proteinase K, but survived heating, indicating that the inhibitory protein is heat-stable. The inhibitory protein was found to affect the attachment step in biofilm formation, but had no effect on preformed biofilms. Replacement of saeSL with saeSP in the biofilm-proficient S. aureus USA300 FPR3757 resulted in the loss of biofilm formation. Culture supernatants of USA300 FPR3757 saeSP, did not inhibit biofilm formation by other staphylococci, suggesting that the inhibitory factor is produced but not secreted in the mutant strain. A number of biochemical methods were utilized to isolate the inhibitory protein. Although a number of candidate proteins were identified, none were found to be the actual inhibitor. In an effort to reduce the number of potential inhibitory genes, RNA-Seq analyses were done with wild-type strain Newman and the saeSL and ΔsaeRS mutants. RNA-Seq results indicated that sae regulates many genes that may affect biofilm formation by Newman.
Project description:We report the use of Next Generation RNA Sequencing for confirming or improving initial identification of small regulatory RNA in Staphylococcus aureus to prodive a list of the most probable RNA molecules transcribed as independent units. Extraction of RNAs in exponential phase of growth in Newman and N315 strains
Project description:BRcells and DRcells show distinct transcriptomic profiles. We assessed transcriptome changes, by deep-sequencing, of different subpopulations of Staphylococcus aureus strain Newman from multicellular aggregates.
Project description:The S. aureus transcriptome was assessed for strains Newman (wild type) and Newman (sarZ) during both exponential (2hr) and early stationary (5hr) cell growth.
Project description:Staphylococcus aureus Newman and Staphylococcus epidermidis Tu3298, 20 minutes post challenge with sub-inhibitory concentration of sapienic acid vs equivalent concentration of ethanol. Challenge was added at mid logarithmic growth (OD600 0.5). Biological triplicates of samples were sequenced.
