Epigenetic Modification Impacts on Gene Regulation in Streptococcus pneumoniae. Issue 5 (27th May 2022)

• Record Type: Journal Article
• Authors: Althari, Yasmeen; Chen, Yongkui; Haigh, Richard; Oggioni, Marco
• Is Part Of: Access microbiology. Volume 4:Issue 5(2022)
• Publication Date: 2022-05-27
• Subjects: Microbiology -- Periodicals; 579
• DOI: 10.1099/acmi.ac2021.po0136
• Languages: English
• ISSNs: 2516-8290

Abstract

Abstract : Methylation by the type I restriction modification system (RMS) SpnIII in Streptococcus pneumoniae is hypothesised to regulate gene expression via epigenetic changes (Manos). The phase variable SpnIII generates six host specificity determinants (hsdS) alleles through site-specific recombination (DeSteCroix), each allele correspond with different methylation pattern. In addition to known functions of the RMS in restricting phage infection (Furi) and transformation (Kwun), we have now tested impact on gene expression. RNAseq was used to analyse S. pneumoniae strains expressing a single spnIII allele (spnIIIA, spnIIIB or spnIIIE) to determine differences in gene expression profiles. The data have identified six genes which show differential expression and have a methylation site mapping to their predicted promoter region. Three synthetic promoters with the wild type and altered methylation target site were cloned in front of a luciferase gene in strains expressing a single spnIII alleles. For the first three sets of constructs analysed, data indicate that the methylated promoters show a three- to twenty-fold higher activity compared to non-methylated promoters. Results obtained were further confirmed by qPCR analysis. Preliminary data using drugs targeting DNA topoisomerases indicate that the mechanism by which methylation impact gene expression could be related to DNA topology. These data demonstrate for the first-time RMS dependent variations in gene expression and propose an alternative mechanism for this epigenetic regulation. References: * Manso, Nature Communication, 2014; 5:5055. * De Ste Croix, J. Bacteriol. 2019; 201(15). pii: e00233-19. * Furi, J. Bacteriol. 2019; 201(19). pii: e00370-19. * Kwun, Nucleic Acids Res. 2018; 46(21):11438-11453.