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

Global Dynamics of the <i>Escherichia coli</i> Proteome and Phosphoproteome During Growth in Minimal Medium

Recent phosphoproteomics studies have generated relatively large data sets of bacterial proteins phosphorylated on serine, threonine, and tyrosine, implicating this type of phosphorylation in the regulation of vital processes of a bacterial cell; however, most phosphoproteomics studies in bacteria were so far qualitative. Here we applied stable isotope labeling by amino acids in cell culture (SILAC) to perform a quantitative analysis of proteome and phosphoproteome dynamics of Escherichia coli during five distinct phases of growth in the minimal medium. Combining two triple-SILAC experiments, we detected a total of 2118 proteins and quantified relative dynamics of 1984 proteins in all measured phases of growth, including 570 proteins associated with cell wall and membrane. In the phosphoproteomic experiment, we detected 150 Ser/Thr/Tyr phosphorylation events, of which 108 were localized to a specific amino acid residue and 76 were quantified in all phases of growth. Clustering analysis of SILAC ratios revealed distinct sets of coregulated proteins for each analyzed phase of growth and overrepresentation of membrane proteins in transition between exponential and stationary phases. The proteomics data indicated that proteins related to stress response typically associated with the stationary phase, including RpoS-dependent proteins, had increasing levels already during earlier phases of growth. Application of SILAC enabled us to measure median occupancies of phosphorylation sites, which were generally low (<12%). Interestingly, the phosphoproteome analysis showed a global increase of protein phosphorylation levels in the late stationary phase, pointing to a likely role of this modification in later phases of growth

Global Dynamics of the <i>Escherichia coli</i> Proteome and Phosphoproteome During Growth in Minimal Medium

Recent phosphoproteomics studies have generated relatively large data sets of bacterial proteins phosphorylated on serine, threonine, and tyrosine, implicating this type of phosphorylation in the regulation of vital processes of a bacterial cell; however, most phosphoproteomics studies in bacteria were so far qualitative. Here we applied stable isotope labeling by amino acids in cell culture (SILAC) to perform a quantitative analysis of proteome and phosphoproteome dynamics of Escherichia coli during five distinct phases of growth in the minimal medium. Combining two triple-SILAC experiments, we detected a total of 2118 proteins and quantified relative dynamics of 1984 proteins in all measured phases of growth, including 570 proteins associated with cell wall and membrane. In the phosphoproteomic experiment, we detected 150 Ser/Thr/Tyr phosphorylation events, of which 108 were localized to a specific amino acid residue and 76 were quantified in all phases of growth. Clustering analysis of SILAC ratios revealed distinct sets of coregulated proteins for each analyzed phase of growth and overrepresentation of membrane proteins in transition between exponential and stationary phases. The proteomics data indicated that proteins related to stress response typically associated with the stationary phase, including RpoS-dependent proteins, had increasing levels already during earlier phases of growth. Application of SILAC enabled us to measure median occupancies of phosphorylation sites, which were generally low (<12%). Interestingly, the phosphoproteome analysis showed a global increase of protein phosphorylation levels in the late stationary phase, pointing to a likely role of this modification in later phases of growth

Global Dynamics of the <i>Escherichia coli</i> Proteome and Phosphoproteome During Growth in Minimal Medium

Recent phosphoproteomics studies have generated relatively large data sets of bacterial proteins phosphorylated on serine, threonine, and tyrosine, implicating this type of phosphorylation in the regulation of vital processes of a bacterial cell; however, most phosphoproteomics studies in bacteria were so far qualitative. Here we applied stable isotope labeling by amino acids in cell culture (SILAC) to perform a quantitative analysis of proteome and phosphoproteome dynamics of Escherichia coli during five distinct phases of growth in the minimal medium. Combining two triple-SILAC experiments, we detected a total of 2118 proteins and quantified relative dynamics of 1984 proteins in all measured phases of growth, including 570 proteins associated with cell wall and membrane. In the phosphoproteomic experiment, we detected 150 Ser/Thr/Tyr phosphorylation events, of which 108 were localized to a specific amino acid residue and 76 were quantified in all phases of growth. Clustering analysis of SILAC ratios revealed distinct sets of coregulated proteins for each analyzed phase of growth and overrepresentation of membrane proteins in transition between exponential and stationary phases. The proteomics data indicated that proteins related to stress response typically associated with the stationary phase, including RpoS-dependent proteins, had increasing levels already during earlier phases of growth. Application of SILAC enabled us to measure median occupancies of phosphorylation sites, which were generally low (<12%). Interestingly, the phosphoproteome analysis showed a global increase of protein phosphorylation levels in the late stationary phase, pointing to a likely role of this modification in later phases of growth

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 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

Additional file 7: Table S4. of Phosphoproteome dynamics mediate revival of bacterial spores

Estimated occupancies of phosphorylation sites detected in this study. (XLSX 44 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 9: Figure S4. of Phosphoproteome dynamics mediate revival of bacterial spores

SspA structure and conservation across Bacillus species. (A) Multiple sequence alignment of B. subtilis SspA and its homologue proteins from representative Bacillus species. Conserved Ser47 residue is highlighted in solid red and other Ser residues are boxed in purple. The corresponding conservation level and consensus sequence are shown below. The multiple sequence alignment was constructed using Jalview. (B) Ribbon diagram of SspA (aa 12–65) protein (cyan) with bound DNA (pink). Ser47 (red) is located at the tip of the second alpha helix. The N and C protein terminals are absent from the structure. Protein structure was predicted by SWISS-MODEL ( http://swissmodel.expasy.org/ ). (PDF 205 kb