Binding of Campylobacter jejuni FliW Adjacent to the CsrA RNA-Binding Pockets Modulates CsrA Regulatory Activity.
Binding of Campylobacter jejuni FliW Adjacent to the CsrA RNA-Binding Pockets Modulates CsrA Regulatory Activity.
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DOI：
10.3389/fmicb.2020.531596
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发表时间：
2020
影响因子：
5.2
通讯作者：
Thompson SA
中科院分区：
文献类型：
--
作者：
Bogacz M;El Abbar FM;Cox CA;Li J;Fiedler JS;Tran LKH;Tran PMH;Daugherty CL;Blake KH;Wang Z;Azadi P;Thompson SA
空肠弯曲杆菌（Campylobacter jejuni CsrA）是一种mrna结合的转录后调节剂，控制着这一重要病原体的许多代谢和毒力相关特征。大肠杆菌CsrA的活性是通过与小的非编码rna （sRNAs）结合来调节的，而空肠C. CsrA的活性是通过与CsrA拮抗剂flw结合来控制的。在本研究中，我们确定了flw在CsrA上的结合位点。空肠C. CsrA的c端相对于srna结合的CsrA蛋白延伸，其缺失使flw的结合失效。采用细菌双杂交实验评估flw与野生型CsrA及其突变体的相互作用，其中每个氨基酸都单独突变。两个CsrA突变（V51A和N55A）导致FliW结合显著降低。V51A和N55A突变体也显示出CsrA-FliW复合物形成的减少，通过尺寸排除色谱和表面等离子体共振进行评估。这些残基在含有CsrA同源物的细菌物种中高度保守，其活性预计受flw调节。FliW结合的位置紧邻CsrA同型二聚体的两个rna结合位点，这表明FliW与CsrA结合通过空间位阻、静电斥力或改变rna结合位点的整体结构来调节其与mRNA靶标的结合能力。
Campylobacter jejuni CsrA is an mRNA-binding, post-transcriptional regulator that controls many metabolic- and virulence-related characteristics of this important pathogen. In contrast to E. coli CsrA, whose activity is modulated by binding to small non-coding RNAs (sRNAs), C. jejuni CsrA activity is controlled by binding to the CsrA antagonist FliW. In this study, we identified the FliW binding site on CsrA. Deletion of the C-terminus of C. jejuni CsrA, which is extended relative to sRNA-binding CsrA proteins, abrogated FliW binding. Bacterial two-hybrid experiments were used to assess the interaction of FliW with wild-type CsrA and mutants thereof, in which every amino acid was individually mutated. Two CsrA mutations (V51A and N55A) resulted in a significant decrease in FliW binding. The V51A and N55A mutants also showed a decrease in CsrA-FliW complex formation, as assessed by size-exclusion chromatography and surface plasmon resonance. These residues were highly conserved in bacterial species containing CsrA orthologs whose activities are predicted to be regulated by FliW. The location of FliW binding was immediately adjacent to the two RNA-binding sites of the CsrA homodimer, suggesting the model that FliW binding to CsrA modulates its ability to bind to its mRNA targets either by steric hindrance, electrostatic repulsion, or by altering the overall structure of the RNA-binding sites.
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影响因子：
3.6
作者：
Mukherjee S;Yakhnin H;Kysela D;Sokoloski J;Babitzke P;Kearns DB
通讯作者：
Kearns DB
影响因子：
6.4
作者：
Oshiro, R. T.;Rajendren, S.;Kearns, D. B.
通讯作者：
Kearns, D. B.
影响因子：
5.6
作者：
Mercante, Jeffrey;Edwards, Adrianne N.;Romeo, Tony
通讯作者：
Romeo, Tony
DOI：
10.1073/pnas.1602455113
发表时间：
2016-08-30
影响因子：
11.1
作者：
Mukherjee, Sampriti;Oshiro, Reid T.;Kearns, Daniel B.
通讯作者：
Kearns, Daniel B.
影响因子：
16.6
作者：
Dugar G;Svensson SL;Bischler T;Wäldchen S;Reinhardt R;Sauer M;Sharma CM
通讯作者：
Sharma CM