Structural insights into RapZ-mediated regulation of bacterial amino-sugar metabolism.

Structural insights into RapZ-mediated regulation of bacterial amino-sugar metabolism.
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DOI:
10.1093/nar/gkx732
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发表时间:
2017-10-13
影响因子:
14.9
通讯作者:
Luisi BF
Luisi BF
中科院分区:
生物学2区
文献类型:
--
作者:
Gonzalez GM;Durica-Mitic S;Hardwick SW;Moncrieffe MC;Resch M;Neumann P;Ficner R;Görke B;Luisi BF

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在系统发育多样化的细菌中,保守蛋白 RapZ 在 RNA 介导的氨基糖代谢调节中发挥着核心作用。 RapZ 通过选择性地将调节性小 RNA GlmZ 呈递给必需核糖核酸酶 RNase E 使其失活,从而有助于控制磷酸葡萄糖胺的生物发生。在这里,我们报告了全长大肠杆菌 RapZ 的晶体结构(3.40 Å 和 3.25 Å),以及其分离的 C 末端结构域(1.17 Å 分辨率)。结构数据证实,RapZ 的 N 端结构域具有激酶折叠,而 C 端结构域与 6-磷酸果糖激酶(糖酵解途径中的一种重要酶)的亚结构域具有最接近的同源性。 RapZ 自关联成二聚体的结构域交换二聚体,体内数据支持四级结构在 RNA 介导的靶基因表达调控中的重要性。基于生化、结构和遗传数据,我们提出了 GlmZ 的 RapZ 和密切相关的诱饵 sRNA GlmY 的结合和呈递机制。我们讨论了通过重新利用中央代谢中的酶成分来实现 RapZ 分子进化的场景。
In phylogenetically diverse bacteria, the conserved protein RapZ plays a central role in RNA-mediated regulation of amino-sugar metabolism. RapZ contributes to the control of glucosamine phosphate biogenesis by selectively presenting the regulatory small RNA GlmZ to the essential ribonuclease RNase E for inactivation. Here, we report the crystal structures of full length Escherichia coli RapZ at 3.40 Å and 3.25 Å, and its isolated C-terminal domain at 1.17 Å resolution. The structural data confirm that the N-terminal domain of RapZ possesses a kinase fold, whereas the C-terminal domain bears closest homology to a subdomain of 6-phosphofructokinase, an important enzyme in the glycolytic pathway. RapZ self-associates into a domain swapped dimer of dimers, and in vivo data support the importance of quaternary structure in RNA-mediated regulation of target gene expression. Based on biochemical, structural and genetic data, we suggest a mechanism for binding and presentation by RapZ of GlmZ and the closely related decoy sRNA, GlmY. We discuss a scenario for the molecular evolution of RapZ through re-purpose of enzyme components from central metabolism.
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