Crosstalk between the tricarboxylic acid cycle and peptidoglycan synthesis in Caulobacter crescentus through the homeostatic control of α-ketoglutarate.
Crosstalk between the tricarboxylic acid cycle and peptidoglycan synthesis in Caulobacter crescentus through the homeostatic control of α-ketoglutarate.
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DOI：
10.1371/journal.pgen.1006978
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发表时间：
2017-08
期刊：
影响因子：
4.5
通讯作者：
Jacobs-Wagner C
中科院分区：
文献类型：
--
作者：
Irnov I;Wang Z;Jannetty ND;Bustamante JA;Rhee KY;Jacobs-Wagner C
为了实现强大的复制，细菌必须整合细胞代谢和细胞壁生长。虽然这两个过程已得到很好的表征，但它们之间交叉监管的性质和程度尚不清楚。在这里，我们利用经典遗传学、CRISPRi、代谢组学、转录组学和化学互补方法，证明新月柄杆菌中主调节因子 Hfq 的缺失会改变中枢代谢，并以营养依赖性方式导致细胞形状缺陷。我们证明 hfq 缺失突变体中的细胞形态表型可归因于 α-酮戊二酸 (KG) 稳态的破坏。 KG 除了作为三羧酸 (TCA) 循环的关键中间体之外，还是合成肽聚糖（细菌细胞壁的主要成分）所需的酶促反应的副产物。 hfq 缺失突变体中 KG 的积累会干扰肽聚糖合成，导致细胞形态缺陷并增加对肽聚糖靶向抗生素的敏感性。因此，这项工作揭示了 TCA 循环和细胞壁形态发生之间的直接串扰。这种串扰强调了代谢稳态的重要性，不仅可以确保生物合成前体的充分利用，而且可以防止干扰这些中间体作为副产物产生的细胞过程。细菌以其卓越的繁殖能力而闻名，这是我们经常希望控制的特性。为了成功地进行自我复制，细菌细胞必须通过中央代谢产生能量和构建模块，并合成细胞壁材料以复制其形状和大小。细胞复制过程中细胞代谢和细胞壁生长如何整合仍知之甚少。在这项工作中，我们证明了全局调节因子 Hfq 对于维持新月柄杆菌中心代谢物稳态的重要性。具体来说，我们发现，Hfq 缺失导致中心代谢物 α-酮戊二酸的积累会抑制产生细胞壁构件所需的酶促反应。这种细胞壁合成的代谢依赖性扰动导致细胞形态缺陷，并使细胞更容易受到细胞壁靶向抗生素的影响。鉴于中枢代谢和细胞壁生物合成广泛保守，我们的研究结果提出了组合药物设计的新方法。
To achieve robust replication, bacteria must integrate cellular metabolism and cell wall growth. While these two processes have been well characterized, the nature and extent of cross-regulation between them is not well understood. Here, using classical genetics, CRISPRi, metabolomics, transcriptomics and chemical complementation approaches, we show that a loss of the master regulator Hfq in Caulobacter crescentus alters central metabolism and results in cell shape defects in a nutrient-dependent manner. We demonstrate that the cell morphology phenotype in the hfq deletion mutant is attributable to a disruption of α-ketoglutarate (KG) homeostasis. In addition to serving as a key intermediate of the tricarboxylic acid (TCA) cycle, KG is a by-product of an enzymatic reaction required for the synthesis of peptidoglycan, a major component of the bacterial cell wall. Accumulation of KG in the hfq deletion mutant interferes with peptidoglycan synthesis, resulting in cell morphology defects and increased susceptibility to peptidoglycan-targeting antibiotics. This work thus reveals a direct crosstalk between the TCA cycle and cell wall morphogenesis. This crosstalk highlights the importance of metabolic homeostasis in not only ensuring adequate availability of biosynthetic precursors, but also in preventing interference with cellular processes in which these intermediates arise as by-products. Bacteria are well-known for their remarkable ability to multiply, a property that we often aim to control. To successfully self-replicate, bacterial cells must generate energy and building blocks through central metabolism and synthesize cell wall material to reproduce their shape and size. How cellular metabolism and cell wall growth are integrated during cellular replication remains poorly understood. In this work, we demonstrate the importance of the global regulator Hfq for maintaining the homeostasis of central metabolites in Caulobacter crescentus. Specifically, we show that accumulation of central metabolite α-ketoglutarate caused by the loss of Hfq inhibits an enzymatic reaction needed to produce cell wall building blocks. This metabolism-dependent perturbation of cell wall synthesis results in cell morphological defects and renders the cell more susceptible to cell wall-targeting antibiotics. Given that central metabolism and cell wall biosynthesis are broadly conserved, our findings suggest a new approach for combinatorial drug design.
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DOI：
10.1093/bioinformatics/btu638
发表时间：
2015-01-15
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Anders S;Pyl PT;Huber W
通讯作者：
Huber W
影响因子：
14.9
作者：
Caspi R;Billington R;Ferrer L;Foerster H;Fulcher CA;Keseler IM;Kothari A;Krummenacker M;Latendresse M;Mueller LA;Ong Q;Paley S;Subhraveti P;Weaver DS;Karp PD
通讯作者：
Karp PD
影响因子：
9.9
作者：
通讯作者：
--
影响因子：
3.6
作者：
Dodsworth, JA;Cady, NC;Leigh, JA
通讯作者：
Leigh, JA
影响因子：
3.2
作者：
Boutte, Cara C.;Henry, Jonathan T.;Crosson, Sean
通讯作者：
Crosson, Sean