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DNA methylation from a Type I restriction modification system influences gene expression and virulence in Streptococcus pyogenes [PacBio]

ABSTRACT: We used Pacific Biosciences Single Molecule Real-Time sequencing platform to identify modified motifs in an RM deficient strain of Streptococcus pyogenes

ORGANISM(S): Streptococcus pyogenes

PROVIDER: GSE130428 | GEO | 2019/04/30

REPOSITORIES: GEO

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DNA methylation from a Type I restriction modification system influences gene expression and virulence in Streptococcus pyogenes [RNA-seq]

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DNA methylation from a Type I restriction modification system influences gene expression and virulence in Streptococcus pyogenes.

Project description:DNA methylation is pervasive across all domains of life. In bacteria, the presence of N6-methyladenosine (m6A) has been detected among diverse species, yet the contribution of m6A to the regulation of gene expression is unclear in many organisms. Here we investigated the impact of DNA methylation on gene expression and virulence within the human pathogen Streptococcus pyogenes, or Group A Streptococcus. Single Molecule Real-Time sequencing and subsequent methylation analysis identified 412 putative m6A sites throughout the 1.8 Mb genome. Deletion of the Restriction, Specificity, and Methylation gene subunits (ΔRSM strain) of a putative Type I restriction modification system lost all detectable m6A at the recognition sites and failed to prevent transformation with foreign-methylated DNA. RNA-sequencing identified 20 genes out of 1,895 predicted coding regions with significantly different gene expression. All of the differentially expressed genes were down regulated in the ΔRSM strain relative to the parent strain. Importantly, we found that the presence of m6A DNA modifications affected expression of Mga, a master transcriptional regulator for multiple virulence genes, surface adhesins, and immune-evasion factors in S. pyogenes. Using a murine subcutaneous infection model, mice infected with the ΔRSM strain exhibited an enhanced host immune response with larger skin lesions and increased levels of pro-inflammatory cytokines compared to mice infected with the parent or complemented mutant strains, suggesting alterations in m6A methylation influence virulence. Further, we found that the ΔRSM strain showed poor survival within human neutrophils and reduced adherence to human epithelial cells. These results demonstrate that, in addition to restriction of foreign DNA, gram-positive bacteria also use restriction modification systems to regulate the expression of gene networks important for virulence.

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Project description:In Streptococcus pyogenes, mutation of GidA results in avirulence despite the same growth rate as the wild type. To understand the basis of this effect, global transcription profiling was conducted. Keywords: Wild type vs. GidA mutant Streptococcus pyogenes

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DNA methylation from a Type I restriction modification system influences gene expression and virulence in Streptococcus pyogenes

Project description:This SuperSeries is composed of the SubSeries listed below.

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DNA methylation from a Type I restriction modification system influences gene expression and virulence in Streptococcus pyogenes [PacBio]

Project description:DNA methylation from a Type I restriction modification system influences gene expression and virulence in Streptococcus pyogenes [PacBio]

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Project description:Transcriptional profiling of Streptococcus pyogenes MGAS5005 cells comparing control untreated GAS cells with GAS cells exposed to 4uM heme for 1.5 h

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