Group A Streptococcus Transcriptome Analysis

Sanson, Misu A.

doi:10.1007/978-1-0716-0467-0_8

Cited by 6 publications

(4 citation statements)

References 20 publications

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“…High-quality RNA was extracted from GAS cells grown to mid-exponential phase (optical density at 600 nm [OD 600 ] of 0.5) in THY at 37°C and 5% CO 2 using the Qiagen RNeasy minikit as previously described ( 73 ). The quantity and quality of RNA were assessed using nanodrop (ThermoFisher) and by Agilent TapeStation.…”

Section: Methodsmentioning

confidence: 99%

Population Genomics of emm4 Group A Streptococcus Reveals Progressive Replacement with a Hypervirulent Clone in North America

et al. 2021

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Section: Methodsmentioning

confidence: 99%

Population Genomics of emm4 Group A Streptococcus Reveals Progressive Replacement with a Hypervirulent Clone in North America

et al. 2021

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“…For RNA seq analysis, RNA was isolated from four replicate cultures for each strain grown to mid‐exponential phase in THY (OD ~ 0.5) using the RNeasy kit (Qiagen) and processed as previously described (Horstmann et al., 2014 ). RNAseq data analysis was performed as previously described (Sanson & Flores, 2020 ). As the MGAS2221 genome is not publicly available, we used the MGAS5005 genome as the reference genome, which is identical to MGAS2221 (Sumby et al., 2005 ) in gene content.…”

Section: Methodsmentioning

confidence: 99%

Genome‐wide analysis of in vivo CcpA binding with and without its key co‐factor HPr in the major human pathogen group A Streptococcus

DebRoy

Aliaga-Tobar

Galvez

et al. 2020

Molecular Microbiology

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Catabolite control protein A (CcpA) is a master regulator of carbon source utilization and contributes to the virulence of numerous medically important Gram‐positive bacteria. Most functional assessments of CcpA, including interaction with its key co‐factor HPr, have been performed in nonpathogenic bacteria. In this study we aimed to identify the in vivo DNA binding profile of CcpA and assess the extent to which HPr is required for CcpA‐mediated regulation and DNA binding in the major human pathogen group A Streptococcus (GAS). Using a combination RNAseq/ChIP‐seq approach, we found that CcpA affects transcript levels of 514 of 1667 GAS genes (31%) whereas direct DNA binding was identified for 105 GAS genes. Three of the directly regulated genes encode the key GAS virulence factors Streptolysin S, PrtS (IL‐8 degrading proteinase), and SpeB (cysteine protease). Mutating CcpA Val301 to Ala (strain 2221‐CcpA‐V301A) abolished interaction between CcpA and HPr and impacted the transcript levels of 205 genes (40%) in the total CcpA regulon. By ChIP‐seq analysis, CcpAV301A bound to DNA from 74% of genes bound by wild‐type CcpA, but generally with lower affinity. These data delineate the direct CcpA regulon and clarify the HPr‐dependent and independent activities of CcpA in a key pathogenic bacterium.

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“…Following on-board adapter trimming and quality control, reads were subsequently mapped to the reference emm3 genome (MGAS10870; Accession number CP067090.1) using CLC Genomics Workbench version 23 (Qiagen). Identification of significantly differentially expressed genes was determined as we have previously described (M. Sanson and Flores 2020). Genes for which the change in expression was >1.5 fold and P-value <0.05 following Bonferroni's correction for multiple comparisons were considered to have a significant difference in expression.…”

Section: Rna-sequencing (Rna-seq) and Qrt-pcr Analysismentioning

confidence: 99%

LiaR-dependent gene expression contributes to antimicrobial responses in group AStreptococcus

Vega,

Sansón-Iglesias,

Mukherjee

et al. 2024

Preprint

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The ability to sense and respond to host defenses is essential for pathogen survival. Some mechanisms involve two-component systems (TCS) that respond to host molecules, such as antimicrobial peptides (AMPs) and activate specific gene regulatory pathways to aid in survival. Alongside TCSs, bacteria coordinate cell division proteins, chaperones, cell wall sortases and secretory translocons at discrete locations within the cytoplasmic membrane, referred to as functional membrane microdomains (FMMs). In Group A Streptococcus (GAS), the FMM or “ExPortal” coordinates protein secretion, cell wall synthesis and sensing of AMP-mediated cell envelope stress via the LiaFSR three-component system. Previously we showed GAS exposure to a subset of AMPs (a-defensins) activates the LiaFSR system by disrupting LiaF and LiaS co-localization in the ExPortal, leading to increased LiaR phosphorylation, expression of the transcriptional regulator SpxA2, and altered GAS virulence gene expression. The mechanisms by which LiaFSR integrates cell envelope stress with responses to AMP activity and virulence are not fully elucidated. Here, we show the LiaFSR regulon is comprised of genes encoding SpxA2 and three membrane-associated proteins: a PspC domain-containing protein (PCP), the lipoteichoic acid-modifying protein LafB and the membrane protein insertase YidC2. Our data show phosphorylated LiaR induces transcription of these genes via a conserved operator, whose disruption attenuates GAS virulence and increases susceptibility to AMPs in a manner primarily dependent on differential expression of SpxA2. Our work expands understanding of the LiaFSR regulatory network in GAS and identifies targets for further investigation of mechanisms of cell envelope stress tolerance contributing to GAS pathogenesis.

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Group A Streptococcus Transcriptome Analysis

Cited by 6 publications

References 20 publications

Population Genomics of emm4 Group A Streptococcus Reveals Progressive Replacement with a Hypervirulent Clone in North America

Population Genomics of emm4 Group A Streptococcus Reveals Progressive Replacement with a Hypervirulent Clone in North America

Genome‐wide analysis of in vivo CcpA binding with and without its key co‐factor HPr in the major human pathogen group A Streptococcus

LiaR-dependent gene expression contributes to antimicrobial responses in group AStreptococcus

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