Function of c-di-AMP in Archaea

c-di-AMP 在古细菌中的功能

• 批准号: 423460053
• 负责人: Professorin Dr. Sonja Verena Albers
• 金额: --
• 依托单位: Forschungsschwerpunkt Mikrobiologie
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2019
• 资助国家: 德国
• 起止时间: 2018-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/423460053?language=en
• 关键词: Function; di; AMP; Archaea

Cyclic nucleotides and their role as second messengers for intracellular signal transduction has been well described in bacteria. Recently cyclic di-AMP was discovered as bacterial second messenger, which has been demonstrated to be essential under normal growth conditions in bacteria. Compared to bacteria, very little is known about second messengers in archaea. We have demonstrated that c-di-AMP is present in the extremely halophilic eurayarchaeon Haloferax volcanii. Its genome encodes one diadenylate cyclase (DacZ) which we expressed and purified and demonstrated to produce c-di-AMP in vitro. Like in bacteria, the dacZ gene is essential in H. volcanii under normal growth conditions and overexpression is lethal. This indicates that c-di-AMP levels have to be tightly regulated in H. volcanii. Experiments with an H. volcanii mutant expressing lower levels of the dacZ gene showed that this mutant had an increased cell volume at low salt concentrations, suggesting that c-di-AMP is important for the regulation osmohomeostasis. To understand the role of c-di-AMP in H. volcanii, we will (A) characterize DacZ further and identify the role of the N-terminal pyruvate kinase effector binding domain, (B) characterize the role of c-di-AMP in the regulation of osmohomeostasis in H. volcanii and (C) identify c-di-AMP binding proteins and other proteins that are involved in the c-di-AMP regulatory network in this organism.

环核苷酸及其作为细胞内信号转导的第二信使的作用在细菌中已经被很好地描述。最近发现环二AMP是细菌的第二信使，已被证明在细菌的正常生长条件下是必需的。与细菌相比，人们对古细菌中的第二信使知之甚少。我们已经证明，c-di-AMP是存在于极端嗜盐的eurayarchaeon Haloferax volcanii。它的基因组编码一个二腺苷酸环化酶（DacZ），我们表达和纯化，并证明在体外产生c-di-AMP。和细菌一样，dacZ基因在H. volcano ii在正常生长条件下和过表达是致命的。这表明在H.火山用H做实验。表达较低水平dacZ基因的volcanii突变体表明，该突变体在低盐浓度下细胞体积增加，这表明c-di-AMP对于调节渗透压稳态很重要。了解c-di-AMP在H. volcanii的研究中，我们将（A）进一步表征DacZ并鉴定N-末端丙酮酸激酶效应子结合结构域的作用，（B）表征c-di-AMP在H. volcanii和（C）鉴定了c-di-AMP结合蛋白和参与该生物体中c-di-AMP调节网络的其它蛋白。