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Specialized and shared functions of diguanylate cyclases and phosphodiesterases in Streptomyces development.
Molecular Microbiology ( IF 2.6 ) Pub Date : 2020-08-14 , DOI: 10.1111/mmi.14581
Julian Haist 1 , Sara Alina Neumann 1 , Mahmoud M Al-Bassam 2 , Sandra Lindenberg 1 , Marie A Elliot 3 , Natalia Tschowri 1
Affiliation  

The second messenger bis‐3,5‐cyclic di‐guanosine monophosphate (c‐di‐GMP) determines when Streptomyces initiate sporulation. c‐di‐GMP signals are integrated into the genetic differentiation network by the regulator BldD and the sigma factor σWhiG. However, functions of the development‐specific diguanylate cyclases (DGCs) CdgB and CdgC, and the c‐di‐GMP phosphodiesterases (PDEs) RmdA and RmdB, are poorly understood. Here, we provide biochemical evidence that the GGDEF‐EAL domain protein RmdB from S. venezuelae is a monofunctional PDE that hydrolyzes c‐di‐GMP to 5′pGpG. Despite having an equivalent GGDEF‐EAL domain arrangement, RmdA cleaves c‐di‐GMP to GMP and exhibits residual DGC activity. We show that an intact EAL motif is crucial for the in vivo function of both enzymes since strains expressing protein variants with an AAA motif instead of EAL are delayed in development, similar to null mutants. Transcriptome analysis of ∆cdgB, ∆cdgC, ∆rmdA, and ∆rmdB strains revealed that the c‐di‐GMP specified by these enzymes has a global regulatory role, with about 20% of all S. venezuelae genes being differentially expressed in the cdgC mutant. Our data suggest that the major c‐di‐GMP‐controlled targets determining the timing and mode of sporulation are genes involved in cell division and the production of the hydrophobic sheath that covers Streptomyces aerial hyphae and spores.

中文翻译:

二鸟苷酸环化酶和磷酸二酯酶在链霉菌发育中的特殊和共享功能。

第二信使双 3,5-环二鸟苷单磷酸 (c-di-GMP) 决定链霉菌何时开始孢子形成。c-di-GMP 信号通过调节器 BldD 和西格玛因子 σ WhiG整合到遗传分化网络中。然而,发育特异性双鸟苷酸环化酶 (DGC) CdgB 和 CdgC 以及 c-di-GMP 磷酸二酯酶 (PDE) RmdA 和 RmdB 的功能知之甚少。在这里,我们提供了来自S. venezuelae的 GGDEF-EAL 结构域蛋白 RmdB 的生化证据是一种单功能 PDE,可将 c-di-GMP 水解为 5'pGpG。尽管具有等效的 GGDEF-EAL 结构域排列,但 RmdA 将 c-di-GMP 切割为 GMP 并表现出残留的 DGC 活性。我们表明完整的 EAL 基序对于两种酶的体内功能至关重要,因为表达具有 AAA 基序而不是 EAL 的蛋白质变体的菌株在发育中被延迟,类似于空突变体。∆ cdgB、∆ cdgC、∆ rmdA和 ∆ rmdB菌株的转录组分析表明,这些酶指定的 c-di-GMP 具有全球调控作用,大约 20% 的委内瑞拉链球菌基因在cdgC 中差异表达突变体。我们的数据表明,决定孢子形成时间和模式的主要 c-di-GMP 控制目标是参与细胞分裂和产生覆盖链霉菌气生菌丝和孢子的疏水鞘的基因。
更新日期:2020-08-14
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