Characterization and analysis of a novel diguanylate cyclase PA0847 from Pseudomonas aeruginosa PAO1
Characterization and analysis of a novel diguanylate cyclase PA0847 from Pseudomonas aeruginosa PAO1
复制标题
铜绿假单胞菌 PAO1 中新型二鸟苷酸环化酶 PA0847 的表征和分析
DOI：
10.2147/idr.s194462
复制
发表时间：
2019-03
影响因子：
3.9
通讯作者：
Huang Weidong
中科院分区：
文献类型：
--
作者：
Zhang Yan;Guo Jiayi;Zhang Ning;Yuan Wensu;Lin Zhi;Huang Weidong
背景：环状二鸟苷(c-di-GMP)作为一种中枢信号分子，被发现对多种细菌表型具有调控作用，尤其是与病原菌感染和耐药有关的细菌表型。值得注意的是，许多微生物有多达数十种参与c-di-GMP新陈代谢的蛋白质。这种明显的冗余性和相关的功能特异性成为研究的重点。虽然这些蛋白中有许多已经被鉴定和研究，但来自铜绿假单胞菌PAO1的一种含有PAS和GGDEF结构域的蛋白PA0847的功能仍然不清楚。材料和方法：本研究应用微生物学、生物化学和结构生物学方法对PA0847的基因/蛋白进行了研究。结果：PA0847对细菌的运动能力有影响，但对生物膜的形成无影响。我们记录了氨基酸和化合物对表型的影响，发现PA0847参与了对各种环境营养物质和因素的响应，表明它可能在感知环境线索方面发挥作用。体外和体内研究表明，PA0847是一种活性较强的二鸟苷环化酶(DGC)，其活性依赖于邻近的PAS结构域。有趣的是，PA0847没有显示出显著的产物抑制作用，尽管c-di-GMP结合的两个I位点的关键残基在其GGDEF结构域是保守的。由相邻酪氨酸残基引起的局部结构变化表明了GGDEF家族蛋白的结构和功能的多样性。结论：我们的研究结果为了解c-di-GMP代谢蛋白PA0847的信号机制提供了依据。
Background: As a central signaling molecule, cyclic diguanylate (c-di-GMP) is found to regulate various bacterial phenotypes, especially those involved in pathogen infection and drug resistance. Noticeably, many microbes have up to dozens of proteins that are involved in c-di-GMP metabolism. This apparent redundancy and the relevant functional specificity have become the focus of research. While a number of these proteins have been identified and investigated, the functions of PA0847, a PAS and GGDEF domain-containing protein from Pseudomonas aeruginosa PAO1, remain unclear. Materials and methods: In the current study, microbiology, biochemistry and structural biology methods were applied to characterize the gene/protein of PA0847. Results: We showed that PA0847 affects bacterial motility but not biofilm formation. We recorded the phenotypic influences of amino acids and compounds, and found that PA0847 is involved in response to various environmental nutrients and factors, suggesting its possible role in sensing environmental cues. Both in-vitro and in-vivo studies showed that PA0847 is an active diguanylate cyclase (DGC), whose activity depends on the neighboring PAS domain. Interestingly, PA0847 demonstrates no significant product inhibition, though the key residues of two I-sites for c-di-GMP binding are conserved in its GGDEF domain. A local structural change imposed by an adjacent tyrosine residue was identified, which indicates the structural and functional diversities of the GGDEF family proteins. Conclusion: Our data provide evidence for understanding the signaling mechanism of the unique c-di-GMP metabolizing protein PA0847.
登录
查看更多内容
影响因子：
4.6
作者：
Rossello J;Lima A;Gil M;Rodríguez Duarte J;Correa A;Carvalho PC;Kierbel A;Durán R
通讯作者：
Durán R
二甘氨酸酸环化酶反应调节剂WSPR激活和自抑制的决定因素。
影响因子：
5.6
作者：
De, Nabanita;Navarro, Marcos V. A. S.;Raghavan, Rahul V.;Sondermann, Holger
通讯作者：
Sondermann, Holger
SuhB 通过 Gac/Rsm 途径和 c-di-GMP 信号调节铜绿假单胞菌中的运动-固着开关
影响因子：
5.2
作者：
Li K;Yang G;Debru AB;Li P;Zong L;Li P;Xu T;Wu W;Jin S;Bao Q
通讯作者：
Bao Q
影响因子：
4.2
作者：
Galperin MY
通讯作者：
Galperin MY
影响因子：
2.6
作者：
Bernier, Steve P.;Ha, Dae-Gon;Khan, Wajiha;Merritt, Judith H.;O'Toole, George A.
通讯作者：
O'Toole, George A.