Protein kinase B controls Mycobacterium tuberculosis growth via phosphorylation of the global transcriptional regulator Lsr2
Protein kinase B controls Mycobacterium tuberculosis growth via phosphorylation of the global transcriptional regulator Lsr2
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蛋白激酶 B 通过全局转录调节因子 Lsr2 的磷酸化来控制结核分枝杆菌的生长
DOI：
10.1101/571406
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Alqaseer K
中科院分区：
文献类型：
--
作者：
Alqaseer K
结核分枝杆菌（Mtb）能够在体内持续数月的多种药物治疗。分枝杆菌具有广泛的调节蛋白，包括在生长期间控制肽聚糖生物合成的蛋白激酶B（PknB）。在这里，我们观察到PknB的耗尽导致特异性转录变化，这可能是由H-NS样调节因子Lsr 2在苏氨酸112处的磷酸化减少引起的。PknB对该磷酸化位点的活性用纯化的蛋白质证实，并且该位点是Mtb适应缺氧条件和在固体培养基上生长所必需的。像H-NS一样，Lsr 2以序列依赖性和非特异性模式结合DNA。通过荧光各向异性和电泳迁移率变化测定，Lsr 2的PknB磷酸化降低了DNA结合，我们的磷酸化模拟T112 D Lsr 2的NMR结构表明，这可能是由于DNA结合结构域的动力学增加。相反，磷酸化T112 A Lsr 2在ChIP测序中增加了与某些DNA位点的结合，含有该变体的Mtb显示出与DNA结合变化相对应的转录变化。总之，PknB通过磷酸化全局转录调节因子Lsr 2来控制Mtb的生长和对不断变化的宿主环境的适应。
Mycobacterium tuberculosis(Mtb)is able to persist in the body through months of multi‐drug therapy. Mycobacteria possess a wide range of regulatory proteins, including the protein kinase B (PknB) which controls peptidoglycan biosynthesis during growth. Here, we observed that depletion of PknB resulted in specific transcriptional changes that are likely caused by reduced phosphorylation of the H‐NS‐like regulator Lsr2 at threonine 112. The activity of PknB towards this phosphosite was confirmed with purified proteins, and this site was required for adaptation ofMtbto hypoxic conditions, and growth on solid media. Like H‐NS, Lsr2 binds DNA in sequence‐dependent and non‐specific modes. PknB phosphorylation of Lsr2 reduced DNA binding, measured by fluorescence anisotropy and electrophoretic mobility shift assays, and our NMR structure of phosphomimetic T112D Lsr2 suggests that this may be due to increased dynamics of the DNA‐binding domain. Conversely, the phosphoablative T112A Lsr2 had increased binding to certain DNA sites in ChIP‐sequencing, andMtbcontaining this variant showed transcriptional changes that correspond with the change in DNA binding. In summary, PknB controlsMtbgrowth and adaptations to the changing host environment by phosphorylating the global transcriptional regulator Lsr2.
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影响因子：
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作者：
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DOI：
10.1073/pnas.1710167114
发表时间：
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影响因子：
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影响因子：
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作者：
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影响因子：
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