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A genome-wide analysis of small regulatory RNAs in the human pathogen group A Streptococcus

ABSTRACT: This SuperSeries is composed of the following subset Series: GSE17767: Transcriptome comparison of Streptococcus pyogenes strain MGAS2221 with that of an isogenic PEL mutant GSE17789: Tiling microarray-based identification of small regulatory RNAs in the group A Streptococcus (GAS) Refer to individual Series

ORGANISM(S): Streptococcus pyogenes

SUBMITTER: Paul Sumby

PROVIDER: E-GEOD-17790 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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A genome-wide analysis of small regulatory RNAs in the human pathogen group A Streptococcus.

Perez Nataly N Treviño Jeanette J Liu Zhuyun Z Ho Siu Chun Michael SC Babitzke Paul P Sumby Paul P

PloS one 20091102 11

The coordinated regulation of gene expression is essential for pathogens to infect and cause disease. A recently appreciated mechanism of regulation is that afforded by small regulatory RNA (sRNA) molecules. Here, we set out to assess the prevalence of sRNAs in the human bacterial pathogen group A Streptococcus (GAS). Genome-wide identification of candidate GAS sRNAs was performed through a tiling Affymetrix microarray approach and identified 40 candidate sRNAs within the M1T1 GAS strain MGAS222 ...[more]

PMID: 19888332

Similar Datasets

Tiling microarray-based identification of small regulatory RNAs in the group A Streptococcus (GAS)

Project description:Small regulatory RNAs (sRNAs) represent a major mechanism of virulence regulation in several pathogens. To date, only three sRNAs have been described in GAS in any detail. To identify whether sRNAs are common within the GAS genome, we created a custom Affymetrix tiling microarray in which all intergenic regions (>49 bp) of the MGAS2221 genome were tiled at high density. Triplicate cultures of strain MGAS2221 in THY broth were grown to the exponential phase (O.D.600 of 0.5) of growth and RNA was isolated, converted to cDNA, fragmented, labeled, and hybridized to our custom array (one array per RNA sample [ hence three arrays total]). Our data identified the presence of 40 candidate sRNAs within the MGAS2221 genome. Three cultures of MGAS2221 were grown in THY broth to the exponential (O.D. 0.5) phase of growth. Two volumes of RNA protect were added to the samples, the samples incubated at room temperature for 5 minutes, and the bacteria collected through centrifugation. Total RNA was isolated via a mechanical disruption method, converted to cDNA, fragmented, labeled, and hybridized to our Affymetrix microarray. Data was analyzed using tiling analysis software to combine the data from the three arrays and generate signal intensities. The supplementary files 'GSE17789_2221sRNA_signal.bar' and 'GSE17789_2221sRNA_pvalue.bar' contain the signals and p-values, respectively, for Samples GSM444010-GSM444012. The BAR files are generated by TAS software using PM/MM signal intensities. Signal intensity data is reported in a linear scale; the p-value is reported in -10log10 scale. Probe analysis used a bandwidth of 40. Interval analysis used a threshold of 0, a maximum gap of 20, and a minimum run of 60.

2010-05-06 | E-GEOD-17789 | biostudies-arrayexpress

Transcriptome comparison of Streptococcus pyogenes strain MGAS2221 with that of an isogenic PEL mutant

Project description:PEL is a small regulatory RNA that is present within the S. pyogenes genome. In some strains of S. pyogenes, the PEL sRNA regulates many of the known virulence factors produced by this pathogen. A genome-wide analysis of PEL-regulated genes had not been performed previously so we analyzed this by comparing the parental M1T1 isolate MGAS2221 with an isogenic PEL mutant. Comparing gene transcript levels between these two strains at the exponential and stationary phases of growth, we identified that PEL has no regulatory function in MGAS2221. Three cultures of each GAS strain were grown in THY broth to exponential (O.D. 0.5) and stationary (O.D. 1.7) phases of growth. Two volumes of RNA protect were added to samples recovered at these O.D.'s, the samples incubated at room temperature for 5 minutes, and the bacteria collected through centrifugation. Total RNA was isolated via a mechanical disruption method, converted to cDNA, fragmented, labeled, and hybridized to our Affymetrix microarray. Estimates of gene expression were calculated using GCOS software v1.4.

2009-08-25 | E-GEOD-17767 | biostudies-arrayexpress

RNAseq analysis of the serotype M1 group A Streptococcus strain MGAS2221

Project description:We recovered RNA from exponential and stationary phase cell cultures of MGAS2221 and utilized two distinct methods to remove rRNA from our samples, the RiboZero kit from Epicenter, and the terminator- 5--Phosphate-Dependent Exonuclease (TEX). This gave us 4 distinct RNAs with which we performed RNAseq analysis. Both rRNA removal methods worked well, with only a small percentage of the final total read counts being derived from rRNA sequences. A single group A Streptococcus strain was investigated.

2013-03-20 | E-GEOD-45325 | biostudies-arrayexpress

Transcriptome comparison of S. pyogenes strains MGAS2221 and 2221ΔFasX following exposure to human plasma

Project description:The fasX gene encodes a small regulatory RNA in the group A Streptococcus (GAS). To determine the FasX regulon during GAS exposure to human plasma we compared parental strain MGAS2221 with isogenic fasX mutant strain 2221ΔFasX. Gene expression was analyzed 0, 15, and 60 minutes post-plasma exposure. GAS strains were grown in THY broth to mid-exponential phase, corresponding to an O.D.600 of 0.5. Two 16ml aliquots of each GAS culture were pelleted by centrifugation and the supernatant removed. Each GAS cell pellet was resuspended in 16ml of pre-warmed human plasma. Thus, four GAS-plasma suspensions were created, two each for each of the GAS strains, with one of the two using plasma from person A, and the other two using plasma from person B. Aliquots were recovered from each of the four GAS-plasma suspensions at T = 0, 15, and 60 mins. For the first two time-points 4.5ml of GAS-plasma suspension were added to 9ml of RNAprotect (Qiagen Inc.). For the final time-point, 4ml was added to 8ml of RNAprotect. Following addition of RNAprotect, samples were incubated at room temperature for 5 mins before centrifuging. Supernatant was removed and the pellets quick frozen in liquid nitrogen and stored at -80 °C until ready to use. RNA isolation, cDNA synthesis, and use of our custom Affymetrix microarray was as previously described (Perez et al., 2009)

2012-07-31 | E-GEOD-36645 | biostudies-arrayexpress

Effect of CovR polymorphisms on GAS 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. 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.

2011-11-01 | E-GEOD-32106 | biostudies-arrayexpress

Comparison of gene expression in MGAS315 and its isogenc mtsR mutant

Project description:RATIONALE: mtsR is a regulator of genes involved in ion aquisition in S. pyogenes that has been suggested to be involved in development of invasive infections. We wanted to identify mtsR regulon. EXPERIMENT: We compared global gene expression in wild type strain MGAS315 with expression in its isogenc mtsR mutant. Experiment was performed with bacteria in exponential (EXP) and at late exponential phase (Late EXP) grown in THY medium Keywords: strain comparison 1. Three independent cultures of WT or mutant strain bacteria at exponential phase of growth (OD60=0.5, biological replicates) served as source of total RNA. RNA was transcribed into cDNA, fragmented and labeled using standard Affymetrix protocol for prokaryotic target preparation. Hybridization with custom Affymetrix array and scaning was performed using protocol recomended by manufacturer. Hybridizaton values aquired by GCOS (in CHP files) are normalized to global hybridization intesity. 2. Analogous experiment was performed with bacteria grown to OD600=1.2 representing late exponential growth phase

2010-05-06 | E-GEOD-10252 | biostudies-arrayexpress

Genome-wide localisation map of ScRad9-9Myc overexpressed in pYX142 grown in SC (synthetic complete medium) along with BCS and BPS copper/iron chelators

Project description:This experiment was performed in order to assess the specificity of Rad9 binding to S. cerevisiae genome. In another ChIP-chip experiment in SC BCS BPS growth conditions we have found Rad9 present to a significant number of genomic loci and wanted to examine how specific this localisation is. For that we inserted Rad9 gene in the high copy plasmid pYX142 and transformed it in BJ5457 rad9delta yeast cells. Cells were grown under the same conditions and ChIP-chip was performed. By this experiment we aimed to examine quantitavely and qualitatively the Rad9-targets when Rad9 is overexpressed and to compare the findings with previous experiments of ours.

2014-11-01 | E-MTAB-2200 | biostudies-arrayexpress

ScRad9-13Myc genome-wide localisation map under galactose induction

Project description:This experiment was performed in order to assess the specificity of Rad9 binding to S. cerevisiae genome. In another ChIP-chip experiment in SC BCS BPS growth conditions we have found Rad9 present to a significant number of genomic loci with a bias to transcriptionally active regions. In order to see if that pattern was random and depended or not on the activity state of the genes, we conducted a Rad9 ChIP on chip experiment with the strain grown in medium with galactose instead of glucose, where it is known that particular gene groups are transcriptionally activated. We then compared to the results of our previous experiment where the strain was grown in glucose.

2014-11-01 | E-MTAB-2201 | biostudies-arrayexpress

Transcriptomes of covR/S mutant strains of S. pyogenes

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.

2009-06-05 | E-GEOD-16436 | biostudies-arrayexpress

Transcriptome comparison of S. pyogenes strains MGAS2221 and 2221?rivRX

Project description:The rivR gene encodes a putative transcriptional regulator, while the rivX gene encodes a putative small regulatory RNA. To determine the RivRX regulon during exponential phase growth in THY broth we compared parental strain MGAS2221 with isogenic rivRX mutant strain 2221?rivRX. GAS strains were grown in THY broth to mid-exponential phase, corresponding to an O.D.600 of 0.5. A 4.5ml aliquot of each GAS culture was added to 9ml of RNAprotect (Qiagen Inc.). Following addition of RNAprotect, samples were incubated at room temperature for 5 mins before centrifuging. Supernatant was removed and the pellets quick frozen in liquid nitrogen and stored at -80 °C until ready to use. RNA isolation, cDNA synthesis, and use of our custom Affymetrix microarray was as previously described (Perez et al., 2009)

2012-12-01 | E-GEOD-36646 | biostudies-arrayexpress

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