Project description:JRS4 is our wild-type M6 serotype strain and JRS948 is the isogenic covR strain. Keywords: 2-channel Four separate RNA preparations from each strain (RNA #A-D) were hybridized to duplicate arrays. Reference RNA consisted of pooled RNA from each preparation, labelled with Cy5.
Project description:JRS4 is our wild-type M6 serotype GAS and JRS519 is our Mga- M6 serotype GAS. SF370 is our wild-type M1 serotype GAS and KSM165L is our Mga- M1 serotype GAS. Table 1: Genomic Normalization Description: Step by Step instructions for the removal of outlier background data and any ORF data that did not meet threshold of 2SD above background average, using original data from gpr file provided in each of the sample files. Table 2: M1 Mga+ vs Mga- Description: Step by Step instructions for the removal of outlier data as compared across all possible data points for each ORF. Data points after genomic normalization are used with resultant averages and standard deviations for each ORF computed. Table 3: M6 Mga+ vs Mga- Description: Step by Step instructions for the removal of outlier data as compared across all possible data points for each ORF. Data points after genomic normalization are used with resultant averages and standard deviations for each ORF computed. Keywords: repeat sample
Project description:In this study we sought to determine the effects of single amino acid replacements in the control of virulence regulatory (CovR) regulatory protein on group A streptococcal global gene expression. We compared the transcriptomes of the serotype M3 strain MGAS10870 with four derivative strains: 1) a strain in which CovR had been completely inactivated (CovR knockout); 2) a strain in which the Arg 144 amino acid was changed to a cysteine (CovR-R144C); 3) a strain in which the Arg 158 amino acid was changed to a cysteine (CovR-R158C); and 4) a strain in which the the Asn 193 amino acid was changed to an isoleucine (CovR-N193I). Duplicate samples of each strain were grown to mid-exponential phase and the relative transcriptomes were compared using an Affymetrix GeneChipl.
Project description:The transcriptional regulator Mga of Streptococcus pyogenes (the group A streptococcus, GAS) is known to directly activate several virulence genes important for colonization and immune evasion during exponential growth. Transcriptome analysis comparing two mga-1 serotypes (M1 SF370, M6 JRS4) and one mga-2 serotype (M4 GA40634) against their isogenic mga-inactivated strains uncovered a broader Mga regulon profile containing both activated and repressed genes with predicted functions primarily related to the uptake and metabolism of sugars. Although the divergent M1 and M4 Mga profiles were similar in size and content, the M6 JRS4 strain was clearly distinct, even from other M6 strains. Real-time RT-PCR and northern blot analysis validated our microarray results and confirmed that established core Mga regulon genes directly activated by Mga (emm, scpA, sof, fba) exhibited the highest activation levels across all strains tested. A novel ORF (Spy2036) encoding a cytosolic hypothetical protein was highly activated in all three serotypes and was called gene regulated by Mga or grm. Mga was shown to bind directly to Pgrm, which overlaps the Mga-regulated Psof in OF+ strains, suggesting that grm is part of the core Mga regulon and is able to activate two divergently transcribed genes from a single site in a class II background. Both class and serotype specific Mga-regulated genes, such as speB, were apparent. In fact, Mga activated speB as long as it was expressed in the wild type strain, although direct binding of Mga to the PspeB promoter could not be demonstrated. Thus, Mga is able to both directly and indirectly regulate genes shown to be important for virulence and the metabolic homeostasis of GAS. Keywords: Wild-type vs Mga-
Project description:Global regulatory roles of the TCS response regulator CovR in GBS serotype 1a A909 Keywords: Strain comparison; global regulation Deletion mutant was examined for alterations in gene expression
Project description:Two covR mutant derivatives of parental strain MGAS2221 were recovered from mice experimentally infected with MGAS2221 and shown to differ in terms of the number and concentration of secreted proteins. One of the covR mutant strains had a secretion phenotype identical to a covS mutant strain, while the other had a secretion phenotype identical to a constructed covR mutant strain. To further investigate the potential differences between the two covR mutant strains we performed expression microarray analysis. Single cultures of each of the four GAS strains tested were grown in THY broth to early exponential phase (O.D. 0.2). Two volumes of RNA protect were added, the samples incubated at room temperature for 5 minutes, and the bacteria collected through centrifugation. Total RNA was isolated via a mechaniscal disruption method, converted to cDNA, fragmented, labeled, and hybridized to our Affymetrix microarray. Estimates of gene expression were calculated using GCOS software v1.4. Data represent probes for serotype M1.
Project description:Global regulatory roles of the TCS response regulator CovR in GBS serotype 1a A909 Keywords: Strain comparison; global regulation Deletion mutant was examined for alterations in gene expression
Project description:In this study we sought to determine the effects of single amino acid replacements in the control of virulence regulatory (CovR) regulatory protein on group A streptococcal global gene expression.