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Series GSE158830 Query DataSets for GSE158830
Status Public on Sep 14, 2021
Title RNase III-CLASH of multi-drug resistant Staphylococcus aureus reveals a regulatory mRNA 3’UTR required for intermediate vancomycin resistance
Organisms Staphylococcus aureus subsp. aureus str. JKD6008; Staphylococcus aureus subsp. aureus str. JKD6009
Experiment type Expression profiling by high throughput sequencing
Summary Treatment of methicillin-resistant Staphylococcus aureus (MRSA) infections is dependant on the efficacy of last-line antibiotics like vancomycin. Vancomycin treatment failure is most commonly linked to the emergence of vancomycin-intermediate resistance in clincal isolates (termed VISA). These isolates have not acquired resistance genes but appear to accumulate a heterogenous collection of single nucleotide polymerisms that collectively alter the physiology of the cell to increase vancomycin tolerance. Cell wall thickening is common among VISA isolates and is thought to decrease vancomycin permeability. Changes in regulatory sRNA expression have been correlated with antibiotic stress responses in VISA isolates however the functions of the vast majority of these RNA regulators is unknown. Here we have used the endoribonuclease RNase III to capture RNA-RNA interactions using an RNA proximity-dependant ligation technique termed CLASH. RNase III-CLASH uncovered hundreds of sRNA-mRNA interactions in vivo allowing functional characterisation of many sRNAs for the first time. Surprisingly, we find that an mRNA encoding an unusually long 3’UTR (here termed vigR) functions as a regulatory ‘hub’ within our RNA-RNA interaction network. We demonstrate vigR promotes expression of the cell wall lytic transglycosylase encoded by isaA through a direct mRNA-mRNA interaction. Further, we find that the vigR mRNA 3’UTR is required for cell wall thickening and that deletion of the vigR 3’UTR re-sensitises VISA to vancomycin. Our results demonstrate the utility of RNase III-CLASH for identifying new regulatory RNA functions and indicate that S. aureus may use mRNA-mRNA interactions to co-ordinate gene expression much more widely than previously appreciated.
 
Overall design This series consists of four different sequencing datasets: 1. RNase III-CLASH (RNA proximity-dependant ligation) data from S. aureus JKD6009, 2. dRNA-seq mapping of RNA 5' end, 3. Term-seq mapping of RNA 3' ends, and 4. Total RNA-sequencing data

Please note that the following update has been made on Jan 2022:
GSM4811620-GSM4811625 raw data files have been replaced (the submitters found that the strains used to generate these FASTQ files had a number of SNPs that might affect the data and generated new sequencing data from re-generated strains).
The previous 'd275v_WTv_DESeq2.xls' was replaced with '2021.03.16_MUTvsWT.DEseq2_combined_output.xlsx'
Three additional samples were included
 
Contributor(s) Mediati D, Wong J, Wu S, Pang I, Tree J
Citation(s) 34372700, 35732665
Submission date Sep 30, 2020
Last update date Jun 30, 2022
Contact name Jai Justin Tree
E-mail(s) j.tree@unsw.edu.au
Phone +61 2 938 59142
Organization name University of New South Wales
Department School of Biotechnology and Biomolecular Sciences
Lab Tree lab
Street address Rm s110 Bldg F25, UNSW, Gate 11 Botany St
City Sydney
State/province NSW
ZIP/Postal code 2033
Country Australia
 
Platforms (3)
GPL29205 Illumina NextSeq 500 (Staphylococcus aureus subsp. aureus str. JKD6009)
GPL29206 Illumina NovaSeq 6000 (Staphylococcus aureus subsp. aureus str. JKD6009)
GPL31256 Illumina NovaSeq 6000 (Staphylococcus aureus subsp. aureus str. JKD6008)
Samples (39)
GSM4811608 JKD6009 rnc-HTF RNase III-CLASH replicate 1
GSM4811609 JKD6009 rnc-HTF RNase III-CLASH replicate 2
GSM4811610 JKD6009 rnc-HTF RNase III-CLASH replicate 3
Relations
BioProject PRJNA666667
SRA SRP285933

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SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE158830_2021.03.16_MUTvsWT.DEseq2_combined_output.xlsx 406.6 Kb (ftp)(http) XLSX
GSE158830_CRISPRi_275_vs_pSD_DESeq2.xls.gz 255.6 Kb (ftp)(http) XLS
GSE158830_DEseq_output_vigR_MutvsREPAIR.xlsx 5.5 Mb (ftp)(http) XLSX
GSE158830_MasterTable_GEO.tsv.gz 73.2 Kb (ftp)(http) TSV
GSE158830_RAW.tar 34.2 Mb (http)(custom) TAR (of BEDGRAPH, GR)
GSE158830_delta_275_vs_WT_DESeq2.xls.gz 302.0 Kb (ftp)(http) XLS
GSE158830_pSD275v_pSDv_DESeq2.xls.gz 154.0 Kb (ftp)(http) XLS
SRA Run SelectorHelp
Processed data provided as supplementary file
Processed data are available on Series record
Raw data are available in SRA

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