Independent Promoter Recognition by TcpP Precedes Cooperative Promoter Activation by TcpP and ToxR.
Independent Promoter Recognition by TcpP Precedes Cooperative Promoter Activation by TcpP and ToxR.
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DOI：
10.1128/mbio.02213-21
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发表时间：
2021-10-26
期刊：
影响因子：
6.4
通讯作者：
DiRita VJ
中科院分区：
文献类型：
--
作者：
Calkins AL;Demey LM;Karslake JD;Donarski ED;Biteen JS;DiRita VJ
霍乱是一种由革兰氏阴性细菌霍乱弧菌引起的肠道疾病。为了到达肠上皮细胞的表面，增殖并引起疾病，霍乱弧菌严格调节毒力因子如霍乱毒素（ctxAB）和毒素共调节菌毛（tcpA-F）的产生。ToxT直接负责调节这些主要毒力因子，而TcpP和ToxR通过刺激toxT表达间接调节毒力因子的产生。TcpP和ToxR是激活toxT表达所需的膜定位转录激活因子（MLTA）。为了更深入地了解MLTA在膜中如何识别启动子DNA，我们使用光活化定位显微镜跟踪了活细胞中单个TcpP-PAmCherry分子的动态，并识别了异质扩散模式。我们的研究结果提供的证据表明，（i）TcpP存在于三个生物物理状态（快速扩散，中间扩散，和缓慢扩散），（ii）TcpP这些不同的扩散状态之间的转换，（iii）TcpP分子在缓慢扩散状态与toxT启动子相互作用，和（iv）ToxR是不是必不可少的TcpP本地化的toxT启动子。这些数据完善了目前的模型之间的协同TcpP和ToxR在刺激toxT的表达，并表明TcpP定位的toxT启动子独立的ToxR。
Cholera is a diarrheal disease caused by the Gram-negative bacterium Vibrio cholerae. To reach the surface of intestinal epithelial cells, proliferate, and cause disease, V. cholerae tightly regulates the production of virulence factors such as cholera toxin (ctxAB) and the toxin-coregulated pilus (tcpA-F). ToxT is directly responsible for regulating these major virulence factors while TcpP and ToxR indirectly regulate virulence factor production by stimulating toxT expression. TcpP and ToxR are membrane-localized transcription activators (MLTAs) required to activate toxT expression. To gain a deeper understanding of how MLTAs identify promoter DNA while in the membrane, we tracked the dynamics of single TcpP-PAmCherry molecules in live cells using photoactivated localization microscopy and identified heterogeneous diffusion patterns. Our results provide evidence that (i) TcpP exists in three biophysical states (fast diffusion, intermediate diffusion, and slow diffusion), (ii) TcpP transitions between these different diffusion states, (iii) TcpP molecules in the slow diffusion state are interacting with the toxT promoter, and (iv) ToxR is not essential for TcpP to localize the toxT promoter. These data refine the current model of cooperativity between TcpP and ToxR in stimulating toxT expression and demonstrate that TcpP locates the toxT promoter independently of ToxR.
影响因子：
3.6
作者：
Gebhard S;Gaballa A;Helmann JD;Cook GM
通讯作者：
Cook GM
影响因子：
16.6
作者：
Lorent JH;Diaz-Rohrer B;Lin X;Spring K;Gorfe AA;Levental KR;Levental I
通讯作者：
Levental I