A crosstalk between c-di-GMP and cAMP in regulating transcription of GcsA, a diguanylate cyclase involved in swimming motility in Pseudomonas putida
A crosstalk between c-di-GMP and cAMP in regulating transcription of GcsA, a diguanylate cyclase involved in swimming motility in Pseudomonas putida
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c-di-GMP 和 cAMP 之间的串扰调节 GcsA 的转录，GcsA 是一种参与恶臭假单胞菌游泳运动的二鸟苷酸环化酶
DOI：
10.1111/1462-2920.14832
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发表时间：
2019
影响因子：
5.1
通讯作者：
Huang Qiaoyun
中科院分区：
文献类型：
--
作者：
Xiao Yujie;Liu Huizhong;He Meina;Nie Liang;Nie Hailing;Chen Wenli;Huang Qiaoyun
普遍存在的细菌第二信使c‐di‐GMP由二胍酸环化酶（DGC）合成，并由磷酸二酯酶（PDE）降解。恶臭假单胞菌在其基因组中有数十个DGC/PDE编码基因，但这些基因的表型-基因型相关性和转录调控在很大程度上是未知的。在此，我们对aP.putidac - di - GMP -代谢酶GcsA的功能和转录调控进行了表征。GcsA由两个per - ARNT - sim （PAS）结构域组成，随后是一个规范保守中心序列模式（GGDEF）结构域和一个截断的EAL结构域。体外分析证实了GcsA的DGC活性。表型观察显示，GcsA以依赖于FlgZ的方式抑制游泳运动。在转录调控方面，c‐di‐GMP和cAMP分别通过其效应物FleQ和Crp协同调控gcsa2。c‐di‐GMP促进gcsaa的转录，cAMP抑制其转录。玻璃结合分析显示FleQ间接调控gcsa的转录，而Crp直接调控gcsaby与其启动子结合的转录。此外，细胞c - di - GMP与cAMP水平呈反比关系。putidawas证实。这些发现为GcsA的功能和转录调控提供了基础知识，并证明了c - di - GMP和cAMP在GcsA的表达调控中存在串扰。
The ubiquitous bacterial second messenger c‐di‐GMP is synthesized by diguanylate cyclase (DGC) and degraded by phosphodiesterase (PDE).Pseudomonas putidahas dozens of DGC/PDE‐encoding genes in its genome, but the phenotypical–genotypical correlation and transcriptional regulation of these genes are largely unknown. Herein, we characterize function and transcriptional regulation of aP.putidac‐di‐GMP‐metabolizing enzyme, GcsA. GcsA consists of two per‐ARNT‐sim (PAS) domains, followed by a canonical conserved central sequence pattern (GGDEF) domain and a truncated EAL domain.In vitroanalysis confirmed the DGC activity of GcsA. The phenotypic observation revealed that GcsA inhibited swimming motility in an FlgZ‐dependent manner. In terms of transcriptional regulation,gcsAwas found to be cooperatively regulated by c‐di‐GMP and cAMP via their effectors, FleQ and Crp respectively. The transcription ofgcsAwas promoted by c‐di‐GMP and inhibited by cAMP.In vitrobinding analysis revealed that FleQ indirectly regulated the transcription ofgcsA, while Crp directly regulated the transcription ofgcsAby binding to its promoter. Besides, an inverse relationship between the cellular c‐di‐GMP and cAMP levels inP.putidawas confirmed. These findings provide basic knowledge regarding the function and transcriptional regulation of GcsA and demonstrate a crosstalk between c‐di‐GMP and cAMP in the regulation of the expression of GcsA inP.putida.