Presentation_1_Autophosphorylation Mechanism of the Ser/Thr Kinase Stk1 From Staphylococcus aureus.pdf

<p>The eukaryotic-like Ser/Thr kinase Stk1 is crucial for virulence, cell wall biosynthesis, and drug susceptibility in methicillin-resistant Staphylococcus aureus (S. aureus) (MRSA). Importantly, MRSA lacking Stk1 become sensitive to β-lactam antibiotics, implying that Stk1 could be an alternative target for combination therapy. However, the autophosphorylation mechanism of Stk1 remains elusive. Using a phosphoproteomic study, we identified six in vivo phosphorylated activation loop residues (Ser159, Thr161, Ser162, Thr164, Thr166, and Thr172) of Stk1, which are also phosphorylated in vitro. We further showed that cis autophosphorylation of Thr172 in the GT/S motif is essential for self-activation and kinase activity of Stk1 kinase domain (Stk1-KD), whereas the trans autophosphorylation of other activation loop serines/threonines are required for the optimal kinase activity of Stk1-KD. Moreover, substitution of the activation loop serines/threonines impaired in vivo autophosphorylation activity of kinase variants, while T172A and T172D variants were unable to autophosphorylate in the cellular content, underlining the essential role of Thr172 for Stk1 activity in vivo. This study provides insights into molecular basis for regulation of Stk1 activity from S. aureus.</p

Mean clustering coefficients (Cp) and mean absolute path lengths (Lp) in APOE <i>ε</i>4 carriers and APOE <i>ε</i>4 noncarriers.

<p>Mean clustering coefficients (Cp) and mean absolute path lengths (Lp) in APOE <i>ε</i>4 carriers and APOE <i>ε</i>4 noncarriers.</p

Abnormal interregional correlations in APOE <i>ε</i>4 carriers compared with APOE <i>ε</i>4 noncarriers.

<p>The yellow dots indicate the AAL brain regions which showed the significantly abnormal correlations. The red and blue lines indicate the significantly increased and decreased interregional correlations.</p

A. The hub regions in APOE <i>ε</i>4 carriers (The size of the circles represents the nodal centrality in this brain region).

<p>B. The hub regions in APOE <i>ε</i>4 noncarriers. C. Differences of between-group nodal centrality in the APOE <i>ε</i>4 carriers compared with the APOE <i>ε</i>4 noncarriers. The six hub regions which are at least in a network of the APOE <i>ε</i>4 carriers and the APOE <i>ε</i>4 noncarriers and these hub regions indicate the significant between-group differences(p<0.05). The blue spheres represent nodal centrality with significant decreases and the red spheres represent nodal centrality with significant increases in the APOE <i>ε</i>4 carriers compared with the APOE <i>ε</i>4 noncarriers.</p

The left image indicates the between-group differences in clustering coefficients (Cp) and the right image indicates the between-group differences in absolute path lengths (Lp).

<p>The black open circles show the average values in each property and the black lines indicate the 95% confidence intervals of the between-group differences through 1000 permutation tests in each sparsity.</p

The FAQ scores and the average metabolism in left rolandic operculum and left superior parietal gyrus in APOE <i>ε</i>4 carriers.

<p>The FAQ scores and the average metabolism in left rolandic operculum and left superior parietal gyrus in APOE <i>ε</i>4 carriers.</p

Statistical parametric map showing a higher or lower regional cerebral metabolic rate of glucose (FDR corrected p<0.05) in APOE <i>ε</i>4 carriers compared with APOE <i>ε</i>4 noncarriers.

<p>The color bar indicates the t value. The cluster size > = 5.</p

Hub regions in metabolic networks of the APOE <i>ε</i>4 carriers and the APOE <i>ε</i>4 noncarriers listing by the descending order of the APOE <i>ε</i>4 carriers’ normalized betweenness.

<p>Hub regions in metabolic networks of the APOE <i>ε</i>4 carriers and the APOE <i>ε</i>4 noncarriers listing by the descending order of the APOE <i>ε</i>4 carriers’ normalized betweenness.</p

Small-world properties of the metabolic networks.

<p>Above graphs indicate the changes in the Gamma </p><p></p><p></p><p><mi>γ</mi><mo>=</mo></p><p><mi>C</mi><mi>p</mi></p><p><mi>r</mi><mi>e</mi><mi>a</mi><mi>l</mi></p><p></p><mo>/</mo><p><mi>C</mi><mi>p</mi></p><p><mi>r</mi><mi>a</mi><mi>n</mi><mi>d</mi><mi>o</mi><mi>m</mi></p><p></p><p></p><p></p><p></p> and Lambda <p></p><p></p><p><mi>λ</mi><mo>=</mo></p><p><mi>L</mi><mi>p</mi></p><p><mi>r</mi><mi>e</mi><mi>a</mi><mi>l</mi></p><p></p><mo>/</mo><p><mi>L</mi><mi>p</mi></p><p><mi>r</mi><mi>a</mi><mi>n</mi><mi>d</mi><mi>o</mi><mi>m</mi></p><p></p><p></p><p></p><p></p> in APOE <i>ε</i>4 carriers and APOE <i>ε</i>4 noncarriers (sparsity ranging from 7% to 26%).<p></p