Sequestration of a dual function DNA-binding protein by Vibrio cholerae CRP.
Sequestration of a dual function DNA-binding protein by Vibrio cholerae CRP.
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DOI：
10.1073/pnas.2210115119
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发表时间：
2022-11-16
影响因子：
11.1
通讯作者：
中科院分区：
文献类型：
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40年的研究已经确立了环AMP受体蛋白作为转录的直接调节因子。在这里，我们表明，环磷酸腺苷受体蛋白（CRP）与霍乱弧菌内膜。膜缔合被响应于营养缺乏条件的翻译后赖氨酸修饰破坏。膜相关CRP隔离并灭活双功能蛋白肽酶A。我们发现，细胞质肽酶A和CRP激活表达的环二核苷酸磷酸二酯酶，减少生物膜的形成和细胞溶解，并增加趋化性。我们建议CRP和PepA合作优化细胞对营养限制的应激反应。虽然环磷酸腺苷受体蛋白（CRP）调节全球基因转录的机制已被深入研究了几十年，新的发现仍然有待取得。在这里，我们报告说，在快速增长，CRP协会与两个保守的，双功能的DNA结合蛋白肽酶A（PepA）和细胞膜。这些相互作用是不存在的营养有限的生长条件下，由于翻译后修饰的三个赖氨酸在一个单一的CRP面。虽然同时DNA结合是罕见的，从CRP解离的结果增加PepA占用在许多染色体结合位点和数百个基因的差异调节，包括几个编码环二核苷酸磷酸二酯酶。我们发现PepA抑制生物膜形成并激活运动性/趋化性。我们提出了一个模型，其中膜结合CRP干扰PepA DNA结合。在营养限制下，PepA被释放。CRP和游离PepA共同激活了一种转录反应，促使细菌寻找更适宜的环境。这项工作揭示了CRP在隔离调节蛋白中的功能。更广泛地说，它描述了通过代谢调节的转录因子与细胞膜的关联来调节细菌转录组的范例。
Four decades of research have established the cyclic AMP receptor protein as a direct regulator of transcription. Here, we show that cyclic AMP receptor protein (CRP) associates with the Vibrio cholerae inner membrane. Membrane association is disrupted by post-translational lysine modifications in response to nutrient-poor conditions. Membrane-associated CRP sequesters and inactivates the dual-function protein peptidase A. We show that cytoplasmic peptidase A and CRP activate expression of cyclic dinucleotide phosphodiesterases that decrease biofilm formation and cell lysis and increase chemotaxis. We propose that CRP and PepA collaborate to optimize the cellular stress response to nutrient limitation. Although the mechanism by which the cyclic AMP receptor protein (CRP) regulates global gene transcription has been intensively studied for decades, new discoveries remain to be made. Here, we report that, during rapid growth, CRP associates with both the well-conserved, dual-function DNA-binding protein peptidase A (PepA) and the cell membrane. These interactions are not present under nutrient-limited growth conditions, due to post-translational modification of three lysines on a single face of CRP. Although coincident DNA binding is rare, dissociation from CRP results in increased PepA occupancy at many chromosomal binding sites and differential regulation of hundreds of genes, including several encoding cyclic dinucleotide phosphodiesterases. We show that PepA represses biofilm formation and activates motility/chemotaxis. We propose a model in which membrane-bound CRP interferes with PepA DNA binding. Under nutrient limitation, PepA is released. Together, CRP and free PepA activate a transcriptional response that impels the bacterium to seek a more hospitable environment. This work uncovers a function for CRP in the sequestration of a regulatory protein. More broadly, it describes a paradigm of bacterial transcriptome modulation through metabolically regulated association of transcription factors with the cell membrane.
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影响因子：
3.7
作者：
Daubenspeck JM;Liu R;Dybvig K
通讯作者：
Dybvig K
影响因子：
14.9
作者：
EBRIGHT, RH;EBRIGHT, YW;GUNASEKERA, A
通讯作者：
GUNASEKERA, A
影响因子：
5.6
作者：
CHARLIER, D;HASSANZADEH, G;GLANSDORFF, N
通讯作者：
GLANSDORFF, N
影响因子：
3.2
作者：
Devroede, N;Huysveld, N;Charlier, D
通讯作者：
Charlier, D
影响因子：
6.4
作者：
Dalia AB;Lazinski DW;Camilli A
通讯作者：
Camilli A