The FA women's super league : framing developments in elite women's football

In 2009, the Football Association (FA), the national governing body of football in England, announced its plan to introduce the country's first semi-professional women's elite league. Launched in 2010 as the FA Women's Super League (FA WSL), its introduction provided both an opportunity to research whether this evidenced a change of position for the women's elite game within footballing narratives and also to examine the place of the FA within these. This study adopted a critical sociological feminist approach to deconstruct the assumptions, values and practices that frame the female game and the introduction of the FA WSL, while providing new insights into the role of the sport's governing organisation in defining elite women's football. Through observations at matches and interviews with people working within the women's game, an examination of the development and introduction of the FA WSL was undertaken, with valuable early insights provided into the first three years of the new League. The study identified that the introduction of the FA WSL was impacted upon by the complex, closed and gendered nature of the FA's organisational structure. The new League adhered to traditional societal concepts of hegemonic masculinity, heteronormativity and liberal approaches to gender equality. The study also found that the new elite women's structures required the clubs who gained entry into the FA WSL to adhere to commercialised, spectacularised and commodified values which dominate the men's game and neo liberal societal narratives. The increased inclusion of females into elite football structures did not profoundly disrupt traditional discourses or provide evidence of a fundamental challenge to gender inequality in the game

Dynamic Expression of the Translational Machinery during <em>Bacillus subtilis</em> Life Cycle at a Single Cell Level

<div><p>The ability of bacteria to responsively regulate the expression of translation components is crucial for rapid adaptation to fluctuating environments. Utilizing <em>Bacillus subtilis (B. subtilis)</em> as a model organism, we followed the dynamics of the translational machinery at a single cell resolution during growth and differentiation. By comprehensive monitoring the activity of the major <em>rrn</em> promoters and ribosomal protein production, we revealed diverse dynamics between cells grown in rich and poor medium, with the most prominent dissimilarities exhibited during deep stationary phase. Further, the variability pattern of translational activity varied among the cells, being affected by nutrient availability. We have monitored for the first time translational dynamics during the developmental process of sporulation within the two distinct cellular compartments of forespore and mother-cell. Our study uncovers a transient forespore specific increase in expression of translational components. Finally, the contribution of each <em>rrn</em> promoter throughout the bacterium life cycle was found to be relatively constant, implying that differential expression is not the main purpose for the existence of multiple <em>rrn</em> genes. Instead, we propose that coordination of the <em>rrn</em> operons serves as a strategy to rapidly fine tune translational activities in a synchronized fashion to achieve an optimal translation level for a given condition.</p> </div

Expression of ribosomal components is dynamic during vegetative growth.

<p>(<b>A–B</b>) Strains carrying <i>P_rrnA-gfp</i> (AR13) (A-<i>rrnA</i>), <i>P_rrnE-gfp</i> (AR16) (A-<i>rrnE</i>), <i>P_rplA-gfp</i> (AR25) (B-<i>P_rplA</i>), <i>rplA-gfp</i> (AR5) (B-RplA) were grown in rich medium (CH). Samples were taken at the indicated time points [hrs] and the GFP signal monitored using fluorescence microscopy. All fluorescence images were normalized to the same intensity range. Of note, the localization of RplA-GFP was consistent with previous results <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041921#pone.0041921-Lewis1" target="_blank">[78]</a>. Scale bar corresponds to 1 µm. (<b>C</b>) Strains carrying <i>P_rrn-gfp</i> (<i>rrnO, A, B, D, E, I, J</i>), <i>P_rplA-gfp</i> or <i>rplA-gfp</i> were grown in rich medium (CH). Samples were taken at the indicated time points [hrs] and the GFP signal monitored using fluorescence microscopy. The data represent the average of three independent biological repeats. Fluorescence from at least 60 cells was measured and averaged for each time point, and is shown in arbitrary units (a.u.) (see Materials and Methods). (<b>D</b>) Variability of GFP intensity among single cells carrying the indicated <i>P_rrn-gfp</i> reporter at the different time points described in (C). Coefficient of Variability (CV) was calculated as SD divided by mean (see Materials and Methods). (<b>E</b>) Strains carrying <i>P_rrn-gfp</i> (<i>rrnO, A, B, D, E, I, J</i>), <i>P_rplA-gfp</i> or <i>rplA-gfp</i> were grown in minimal (S7) medium and samples processed as in (C). (<b>F</b>) Variability of GFP intensity among single cells carrying the indicated <i>P_rrn-gfp</i> reporter at the different time points described in (E). Coefficient of Variability (CV) was calculated as SD divided by mean (see Materials and Methods).</p

Expression of translational components increases transiently within the forespore.

<p>Strains carrying <i>gltX-gfp</i> (AR11), <i>rnpA-gfp</i> (AR10), <i>thrS-gfp</i> (AR9), <i>rplA-gfp</i> (AR5), or <i>spoIIIG::cat, rplA-gfp</i> (AR20) were induced to sporulate and samples taken at the indicated time points [hrs]. Shown are phase contrast images (left panels) and corresponding GFP fluorescence images (right panels). Arrowheads designate the position of forespores. Scale bar corresponds to 1 µm.</p

Contribution of each rrn promoter to overall translational activity.

<p>The contribution of each <i>rrn</i> promoter during different phases of <i>B. subtilis</i> life cycle is represented as an average percentage, with the sum activity of the seven tested promoters taken as 100%.</p

Bimodal <i>rrn</i> activity during stationary phase.

<p>Cells carrying the P<i>_rrnO-gfp</i> (AR17) were grown in poor medium (S7) (<b>A</b>) or rich medium (CH) (<b>B</b>). Shown are GFP fluorescence images acquired from stationary phase cells (upper panels). Normalized fluorescence distribution of at least 100 individual cells (lower panels) is scored for each phase and is shown in arbitrary units (a.u.). Arrowheads highlight cells displaying either low (A) or high (B) <i>rrn</i> promoter activity, compared to the average level. Scale bars correspond to 1 µm.</p

Additional file 3: Figure S2. of Phosphoproteome dynamics mediate revival of bacterial spores

Comparison of germinating spore and vegetative growth phosphoproteomes. (A) Overlap between the germinating and vegetative phosphoproteins. (B) Common phosphorylation sites between the germinating and vegetative phase phosphoproteins. (C) The identity of the overlapping 17 phosphorylation sites and the stage of vegetative growth at which they show increased phosphorylation. (PDF 185 kb

Additional file 8: Table S5. of Phosphoproteome dynamics mediate revival of bacterial spores

Deletion- and site-directed mutants assessed in this study. (PDF 206 kb

Additional file 16: Figure S9. of Phosphoproteome dynamics mediate revival of bacterial spores

Phospho-modifications of translation elongation factors affect vegetative growth. (A) Spores of PY79 (WT), AR165 (P hyper-spank -EF-G, ∆EF-G), AR166 (P hyper-spank -EF-G-Y339A, ∆EF-G), and AR167 (P hyper-spank -EF-G-Y339D, ∆EF-G) strains were incubated at 37 °C in S7-defined medium supplemented with L-Ala (10 mM) and 0.5 mM IPTG, and optical density (OD600) was measured at the indicated time points. Data are presented as a fraction of the initial OD600 of the phase-bright spores. Decreasing OD600 signifies spore germination while increasing OD600 indicates spore outgrowth. (B) Strains listed in (A) were grown at 37 °C in S7- supplemented with L-Ala (10 mM) and 0.5 mM IPTG, and OD600 was measured at the indicated time points. AR166 (P hyper-spank -EF-G-Y339A, ∆EF-G) and AR167 (P hyper-spank -EF-G-Y339D, ∆EF-G) strains showed significantly reduced growth rates compared to the control strains by repeated measures ANOVA (P <0.05). C) Spores of PY79 (WT), AR157 (P xyl -EF-TU, ∆EF-TU), AR158 (P xyl -EF-TU-Y270A, ∆EF-TU), and AR159 (P xyl -EF-TU-Y270D, ∆EF-TU) strains were incubated at 37 °C in S7- supplemented with L-Ala (10 mM) and 0.5 % xylose, and OD600 was measured at the indicated time points. Data are presented as a fraction of the initial OD600 of the phase-bright spores. (D) Strains listed in (C) were grown at 37 °C in S7- supplemented with L-Ala (10 mM) and 0.5 % xylose, and OD600 was measured at the indicated time points. The data points are averages of results obtained from three independent biological repeats. Error bars designate SD. AR159 (P xyl -EF-TU-Y270D, ∆EF-TU) showed significantly reduced growth rates compared to the other strains by repeated measures ANOVA (P <0.05). (PDF 237 kb

Additional file 15: Figure S8. of Phosphoproteome dynamics mediate revival of bacterial spores

RpsJ structure and conservation across Bacillus species. (A) Multiple sequence alignment of B. subtilis RpsJ with its homologues from representative Bacillus species. Conserved Ser32 residue is highlighted in solid red, the corresponding conservation level and consensus sequence are shown below. The multiple sequence alignment was constructed using Jalview. (B) Ribbon diagram of RpsJ protein Ser32 (red) is located at the tip of globular surface domain. Protein structure was predicted by SWISS-MODEL ( http://swissmodel.expasy.org/ ). (PDF 282 kb