Molecular insights into effector binding by DgoR, a GntR/FadR family transcriptional repressor of D‐galactonate metabolism in Escherichia coli,Molecular Microbiology

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Molecular insights into effector binding by DgoR, a GntR/FadR family transcriptional repressor of D‐galactonate metabolism in Escherichia coli
Molecular Microbiology ( IF 2.6 ) Pub Date : 2020-10-17 , DOI: 10.1111/mmi.14625
Garima Arya ₁ , Mohinder Pal ₁ , Monika Sharma ₂ , Bhupinder Singh ₁ , Swati Singh ₁ , Vishal Agrawal ₃ , Rachna Chaba ₁

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Several GntR/FadR transcriptional regulators govern sugar acid metabolism in bacteria. Although effectors have been identified for a few sugar acid regulators, the mode of effector binding is unknown. Even in the overall FadR subfamily, there are limited details on effector‐regulator interactions. Here, we identified the effector‐binding cavity in Escherichia coli DgoR, a FadR subfamily transcriptional repressor of D‐galactonate metabolism that employs D‐galactonate as its effector. Using a genetic screen, we isolated several dgoR superrepressor alleles. Blind docking suggested eight amino acids corresponding to these alleles to form a part of the effector‐binding cavity. In vivo and in vitro assays showed that these mutations compromise the inducibility of DgoR without affecting its oligomeric status or affinity for target DNA. Taking Bacillus subtilis GntR as a representative, we demonstrated that the effector‐binding cavity is similar among FadR subfamily sugar acid regulators. Finally, a comparison of sugar acid regulators with other FadR members suggested conserved features of effector‐regulator recognition within the FadR subfamily. Sugar acid metabolism is widely implicated in bacterial colonization and virulence. The present study sets the basis to investigate the influence of natural genetic variations in FadR subfamily regulators on their sensitivity to sugar acids and ultimately on host‐bacterial interactions.

中文翻译：

DgoR 对效应子结合的分子见解，DgoR 是大肠杆菌中 D-半乳糖酸代谢的 GntR/FadR 家族转录抑制因子

几个 GntR/FadR 转录调节因子控制细菌中的糖酸代谢。尽管已经确定了一些糖酸调节剂的效应子，但效应子结合的模式尚不清楚。即使在整个 FadR 亚家族中，关于效应器-调节器相互作用的细节也很有限。在这里，我们确定了大肠杆菌DgoR 中的效应子结合腔，这是一种使用 D-半乳糖酸盐作为其效应子的 D-半乳糖酸代谢的 FadR 亚家族转录抑制因子。使用遗传筛选，我们分离了几个dgoR超级阻遏物等位基因。盲对接表明与这些等位基因相对应的八个氨基酸形成效应子结合腔的一部分。体内和体外测定表明，这些突变会损害 DgoR 的诱导性，而不影响其寡聚状态或对靶 DNA 的亲和力。服用枯草芽孢杆菌GntR 作为代表，我们证明了 FadR 亚家族糖酸调节剂之间的效应结合腔相似。最后，糖酸调节剂与其他 FadR 成员的比较表明 FadR 亚家族中效应物-调节剂识别的保守特征。糖酸代谢广泛涉及细菌定植和毒力。本研究为研究 FadR 亚家族调节因子的自然遗传变异对其对糖酸的敏感性以及最终对宿主-细菌相互作用的影响奠定了基础。

更新日期：2020-10-17

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