Lack of the Delta Subunit of RNA Polymerase Increases Virulence Related Traits of Streptococcus mutans

Xue, Xiaoli; Sztajer, Helena; Buddruhs, Nora; Petersen, Jörn; Rohde, Manfred; Talay, Susanne R.; Wagner‐Döbler, Irene

doi:10.1371/journal.pone.0020075

Cited by 19 publications

(21 citation statements)

References 69 publications

(88 reference statements)

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“…However, the utilization of carbon sources could be affected strongly by medium and cultivation conditions. In contrast to the growth condition used in this study (a rich medium and aerobic cultivation with additional CO 2 ), when we used a minimal medium and aerobic cultivation conditions, the mutant showed increased metabolic activity on 20 additional carbon sources in comparison with the wild-type (Xue et al, 2011).…”

Section: Effect Of Rpoe On the S Mutans Proteome Under Various Condimentioning

confidence: 67%

“…an altered cell wall muropeptide pattern and a lower teichoic acid content (Bizzini et al, 2007), reduced capsule expression (Hardy et al, 2000), growth defects (Bizzini et al, 2007;Hardy et al, 2000) and reduced virulence (Bizzini et al, 2007;Buchanan et al, 2005). Thus, the reduction of the level of PGM in the mutant may affect cell wall composition and exopolysaccharide production, which partially explains its altered surface properties and biofilm structure observed in our previous study (Xue et al, 2011).…”

Section: Effect Of Rpoe On the S Mutans Proteome Under Various Condimentioning

confidence: 74%

“…Loss of RpoE caused massive changes in the expression of genes and intergenic regions (Xue et al, 2010). In addition, the mutant had increased virulencerelated traits, including increased adherence to human extracellular matrix components and a broader spectrum of carbon substrates that were respired, and it was resistant to a large number of toxic compounds (Xue et al, 2011). Here we investigated the role of RpoE in the proteome of S.…”

Section: Introductionmentioning

confidence: 99%

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The global impact of the delta subunit RpoE of the RNA polymerase on the proteome of Streptococcus mutans

Xue

Wang

et al. 2012

Microbiology

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Transcriptional specificity in low-G+C Gram-positive bacteria is maintained by RpoE, the delta subunit of the RNA polymerase. Here, we studied the effect of RpoE at the proteome level in the human dental pathogen Streptococcus mutans by comparing the DrpoE mutant with the wild-type under five conditions: (0) exponential growth, (1) early stationary phase, (2) acid stress, (3) oxidative stress, and (4) combined acid and oxidative stress. A total of 280 cellular protein spots were reproducibly detected, of which 97 differentially expressed protein spots were identified by MALDI-TOF MS. Lack of RpoE caused downregulation of proteins for carbohydrate metabolism and energy production, including phosphoglucomutase (PGM), the phosphopentomutase DeoB and the pyruvate formate-lyase Pfl. The DrpoE mutant had extensive changes in the abundance of proteins involved in acid and oxidative tolerance and protein turnover, and of chaperones, at exponential phase in the absence of stress, suggesting a potential internal stress. In addition, the mutant had reduced amounts of proteins for adaptation responses, e.g. the multiple sugar transport and metabolism enzymes required for entering early stationary phase, and the proteins for stress-defence mechanisms and glycolysis under oxidative stress. Comparison of the proteome data with the corresponding transcriptome data suggested that the effects were the result of altered transcriptional and post-transcriptional regulation. The data are consistent with the reduced transcriptional specificity of the RNA polymerase in the DrpoE mutant, and suggest a general impact, but not a specific regulatory role, of RpoE in stress adaptation.

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Section: Effect Of Rpoe On the S Mutans Proteome Under Various Condimentioning

confidence: 67%

Section: Effect Of Rpoe On the S Mutans Proteome Under Various Condimentioning

confidence: 74%

Section: Introductionmentioning

confidence: 99%

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The global impact of the delta subunit RpoE of the RNA polymerase on the proteome of Streptococcus mutans

Xue

Wang

et al. 2012

Microbiology

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“…Growth measurements were performed using PM1-PM8 plates (http://www.biolog.com/pdf/pm_lit/PM1-PM10.pdf) using the standard procedures for E. faecalis (see www.biolog.com and e.g. Xue et al, 2011). The best signal-to-noise ratio was obtained when using pH 5.5 and Biolog Dye H, and these conditions were used throughout.…”

Section: D176/177mentioning

confidence: 99%

EF0176 and EF0177 from Enterococcus faecalis V583 are substrate-binding lipoproteins involved in ABC transporter mediated ribonucleoside uptake

et al. 2015

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One of the ABC transporter systems in Enterococcus faecalis V583 is encoded by the ef0176-ef0180 gene cluster, which differs from orthologous operons in related bacteria in that it contains two genes putatively encoding substrate-binding proteins (SBPs). These SBPs, EF0176 and EF0177, have previously been identified on the surface of E. faecalis. By phenotypic studies of single and double knockout mutants, we show here that EF0176 and EF0177 are specific for ribonucleosides and, by inference, that the EF0176-EF0180 ABC transporter plays a role in nucleoside uptake. The specificity of the SBPs was mapped using growth experiments on a medium, RPMI 1640, that only supports growth of E. faecalis when supplemented with purine nucleosides or their corresponding bases. This analysis was complemented by studies with toxic fluorinated pyrimidine ribonucleoside analogues and competition experiments. The data show that EF0176 and EF0177 have broad and overlapping, but not identical, substrate specificities and that they, together, are likely to bind and facilitate the transport of all common ribonucleosides. Comparative sequence analysis and inspection of an available crystal structure of an orthologue, PnrA from Treponema pallidum, showed that the strongest binding interactions between the protein and the ligand involve the ribose moiety and that sequence variation in the binding site primarily affects interactions with the base. This explains both the broad substrate specificity of these binding proteins and the observed variations therein. The presence of two SBPs in this nucleoside ABC transporter system in E. faecalis may improve the bacterium's ability to scavenge nucleosides.

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“…S. mutans rpoE mutants are also more sensitive to environmental stresses (H 2 O 2 stress and acid stress) and show alterations in biofilm formation and virulence (12). Interestingly, rpoE deletion was also reported to increase self-aggregation, coaggregation with other oral microorganisms (13), and an elevated ability to bind human extracellular matrix proteins. For Streptococcus agalactiae, there have been a limited number of studies which show greatest rpoE expression during exponential growth (14) and attenuated virulence of rpoE mutants in a rat sepsis model (15,16) and human whole-blood survival assays.…”

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confidence: 99%

The δ Subunit of RNA Polymerase Guides Promoter Selectivity and Virulence in Staphylococcus aureus

et al. 2014

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In Gram-positive bacteria, and particularly the Firmicutes, the DNA-dependent RNA polymerase (RNAP) complex contains an additional subunit, termed the ␦ factor, or RpoE. This enigmatic protein has been studied for more than 30 years for various organisms, but its function is still not well understood. In this study, we investigated its role in the major human pathogen Staphylococcus aureus. We showed conservation of important structural regions of RpoE in S. aureus and other species and demonstrated binding to core RNAP that is mediated by the ␤ and/or ␤= subunits. To identify the impact of the ␦ subunit on transcription, we performed transcriptome sequencing (RNA-seq) analysis and observed 191 differentially expressed genes in the rpoE mutant. Ontological analysis revealed, quite strikingly, that many of the downregulated genes were known virulence factors, while several mobile genetic elements (SaPI5 and prophage SA3usa) were strongly upregulated. Phenotypically, the rpoE mutant had decreased accumulation and/or activity of a number of key virulence factors, including alpha toxin, secreted proteases, and Panton-Valentine leukocidin (PVL). We further observed significantly decreased survival of the mutant in whole human blood, increased phagocytosis by human leukocytes, and impaired virulence in a murine model of infection. Collectively, our results demonstrate that the ␦ subunit of RNAP is a critical component of the S. aureus transcription machinery and plays an important role during infection. Bacterial gene transcription is a complex, multifactorial process that involves several key enzymes and regulatory elements. It is driven by the activity of DNA-dependent RNA polymerase (RNAP) and its associated proteins, which form a multisubunit enzyme consisting of one ␤ subunit, one ␤= subunit, two identical ␣ subunits, and one subunit (reviewed in reference 1). Together these form the RNAP apoenzyme, which is able to perform RNA elongation and termination; however, initiation requires the involvement of a factor. Typically, most bacterial species harbor several different factors, a primary one ( A or 70 ) that mediates housekeeping gene transcription and a variety of alternative sigma factors, which aid in the response to unfavorable environmental conditions and stress.In certain Gram-positive species and particularly the Firmicutes (see Fig. S1 in the supplemental material), an additional RNAP subunit is present, termed the ␦ factor, or RpoE (2). In Bacillus subtilis, the 173-amino-acid delta subunit has been shown to reduce nonspecific binding of RNAP to DNA and lead to an elevated preference for DNA regions that include promoter sequences (3, 4). In early experiments, it was shown that RpoE has a role specifically confined to promoter selection and influences the ability of RNAP to form open promoter complexes (5, 6). Fulllength RpoE has also been described as displacing nucleic acids from RNAP-DNA or RNAP-RNA complexes in vitro, which may explain the decreased affinity of RNAP-RpoE for nonpromoter regions. Thi...

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Lack of the Delta Subunit of RNA Polymerase Increases Virulence Related Traits of Streptococcus mutans

Cited by 19 publications

References 69 publications

The global impact of the delta subunit RpoE of the RNA polymerase on the proteome of Streptococcus mutans

The global impact of the delta subunit RpoE of the RNA polymerase on the proteome of Streptococcus mutans

EF0176 and EF0177 from Enterococcus faecalis V583 are substrate-binding lipoproteins involved in ABC transporter mediated ribonucleoside uptake

The δ Subunit of RNA Polymerase Guides Promoter Selectivity and Virulence in Staphylococcus aureus

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