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

Effect of overbalancing phosphorylation activity on the integrity of spore envelopes.

<p>Live-dead staining of spore chains of the eSTPK mutants NLΔPkaI and NLΔ4775–4779 (A) revealed the presence of dead spores (red) or spores without DNA (black). In contrast, spore chains of the parental M145 strain (A) only contained viable spores (green). Expression of a second copy of any eSTPK gene of cluster <i>SCO4775-4779</i> (B-F) caused a similar sporulation defect in <i>S</i>. <i>coelicolor</i> M145, NLΔPkaI, or NLΔ4775–4779. None of the eSTPK genes was able to complement aberrant sporulation of the five-fold mutant NLΔ4775–4779. A, no plasmid integrated; B, :: pSET152-pkaH; C, :: pSET152-SCO4776; D, :: pSET152-pkaD; E, :: pSET152-pkaI; F, :: pSET152-pkaJ. Bar = 5 μm.</p

Effect of overbalancing phosphorylation activity on proper sporulation of <i>S</i>. <i>coelicolor</i>.

<p>Phase contrast microscopy of spore chains revealed the presence of aberrant spores (arrows) in eSTPK mutants NLΔPkaI and NLΔ4775–4779 (A). In contrast, spore chains of the parental M145 strain (A) and the complemented mutant NLΔPkaI::pSET-pkaI (E) contain mainly regular ovoid spores. Not only deletion, but also expression of a second copy of any eSTPK gene of cluster <i>SCO4775-4779</i> causes a similar sporulation defect (B-F, white arrows) in <i>S</i>. <i>coelicolor</i> M145, NLΔPkaI, or NLΔ4775–4779. None of the eSTPK genes is able to complement aberrant sporulation of the five-fold mutant NLΔ4775–4779. A, no plasmid integrated; B, :: pSET152-pkaH; C, :: pSET152-SCO4776; D, :: pSET152-pkaD; E, :: pSET152-pkaI; F, :: pSET152-pkaJ. Bar = 5 μm.</p

Phosphorylation of PBP2 by PkaI.

<p><i>S</i>. <i>coelicolor pbp2</i> with a S-tag encoding sequence was co-expressed with <i>pkaI</i> in <i>E</i>. <i>coli</i>. PBP2_S-tag was purified under denaturing conditions by affinity chromatography. PkaI with an N-terminal His-tag was purified by Ni-NTA chromatography under native conditions. Purified His_PkaI and PBP2_S-tag proteins (bold letters and underlining indicate, which protein was purified) were separated on an SDS polyacrylamide gel and stained with Coomassie blue (<b>A</b>). Phosphorylated proteins were identified by ProQ Diamond staining. The white arrow indicates auto-phosphorylated His_PkaI, while the black arrow marks phosphorylated PBP2_S-tag. Immunoblotting with Anti-S-tag antibodies confirmed the identity of PBP2_S-tag. Domain architecture of PBP2 and positions of the most likely phosphosites (<b>B</b>). Predicted Pfam domains (PBP dimerization, dark grey, PBP transpeptidase, light grey), a transmembrane helix (TM) and the positions of phosphorylated S/T residues, identified by LC-MS/MS, are indicated.</p

Effect of Ser/Thr kinases on the viability of spores (A) and the resistance of germinating spores to vancomycin (B).

<p>Spore chains were stained with the LIVE/DEAD BacLight Bacterial Viability Kit (Molecular Probes) and observed by fluorescence microscopy. Percentage of viable (green), dead (red) and spores without DNA (black) is given for each strain. Spores of the different strains were plated onto LB agar and filter discs containing 5 μg vancomycin were applied. Whereas, M145 and the <i>pkaI</i> mutant NLΔPkaI were resistant, NLΔ4775–4779 spores showed vancomycin sensitivity, suggesting an impaired spore wall. Vancomycin sensitivity of M145 or NLΔPkaI was also caused by expressing a second copy of each kinase gene, with the exception of <i>pkaI</i>. Supplementation of the agar plates with 3 mM MgCl_2, known to rescue mutants impaired in cell wall synthesis restored vancomycin resistance to all strains. A, no plasmid integrated; B, :: pSET152-pkaH; C, :: pSET152-SCO4776; D, :: pSET152-pkaD; E, :: pSET152-pkaI; F, :: pSET152-pkaJ.</p

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 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

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 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 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