BrfA 作为一种细菌增强子结合蛋白,通过感知环状单磷酸二鸟苷来调节荧光假单胞菌中功能性淀粉样蛋白 Fap 依赖性生物膜的形成。

IF 6.1 1区 生物学 Q1 MICROBIOLOGY
Miao Guo , Siqi Tan , Yinying Wu , Chongni Zheng , Peng Du , Junli Zhu , Aihua Sun , Xiaoxiang Liu
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引用次数: 0

摘要

假单胞菌的功能性淀粉样蛋白(Fap)是冷藏海鱼中分离出的荧光假单胞菌 PF07 形成大菌落生物膜、胶粒和固体表面相关(SSA)生物膜的关键。然而,有关 fap 基因表达调控的信息非常有限。在本文中,我们发现一种新型细菌增强子结合蛋白(bEBP)--BrfA--通过直接感知环二鸟苷单磷酸(c-di-GMP)来调控Fap依赖性生物膜的形成。我们的体内数据显示,BrfA的REC结构域缺失促进了fap基因的表达和生物膜的形成,而c-di-GMP以依赖BrfA的方式正向调节fapA的转录。在体外实验中,我们发现 BrfA 的 ATPase 活性受到 REC 结构域的抑制,并被 c-di-GMP 激活。BrfA 和σ因子 RpoN 与 fapA 上游区域结合,BrfA 的结合能力不受 REC 结构域缺失或 c-di-GMP 的影响。BrfA 与 fapA 启动子上游的三个增强子位点特异性结合,这三个位点包含共识序列 CA-(N4)-TGA(A/T)ACACC。使用 lacZ 融合报告基因进行的体内实验表明,所有三个 BrfA 增强子位点都是激活 fapA 转录所必需的。总之,这些发现揭示了 BrfA 是一种新型的 c-di-GMP 响应转录因子,可直接控制荧光团菌中 Fap 生物合成基因的转录。Fap 功能淀粉样蛋白和 BrfA 型转录因子在假单胞菌物种中广泛存在。这项工作提供了有关 c-di-GMP 和 BrfA 依赖性 fap 表达调控的新见解,将有助于开发抗生物膜策略。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
BrfA functions as a bacterial enhancer-binding protein to regulate functional amyloid Fap-dependent biofilm formation in Pseudomonas fluorescens by sensing cyclic diguanosine monophosphate

The functional amyloid of Pseudomonas (Fap) is essential for the formation of macrocolony biofilms, pellicles, and solid surface-associated (SSA) biofilms of Pseudomonas fluorescens PF07, an isolate from refrigerated marine fish. However, limited information on the expression regulation of fap genes is available. Herein, we found that a novel bacterial enhancer-binding protein (bEBP), BrfA, regulated Fap-dependent biofilm formation by directly sensing cyclic diguanosine monophosphate (c-di-GMP). Our in vivo data showed that the REC domain deletion of BrfA promoted fap gene expression and biofilm formation, and c-di-GMP positively regulated the transcription of fapA in a BrfA-dependent manner. In in vitro experiments, we found that the ATPase activity of BrfA was inhibited by the REC domain and was activated by c-di-GMP. BrfA and the sigma factor RpoN bound to the upstream region of fapA, and the binding ability of BrfA was not affected by either deletion of the REC domain or c-di-GMP. BrfA specifically bound to the three enhancer sites upstream of the fapA promoter, which contain the consensus sequence CA-(N4)-TGA(A/T)ACACC. In vivo experiments using a lacZ fusion reporter indicated that all three BrfA enhancer sites were essential for the activation of fapA transcription. Overall, these findings reveal that BrfA is a new type of c-di-GMP-responsive transcription factor that directly controls the transcription of Fap biosynthesis genes in P. fluorescens. Fap functional amyloids and BrfA-type transcription factors are widespread in Pseudomonas species. The novel insights into the c-di-GMP- and BrfA-dependent expression regulation of fap provided by this work will contribute to the development of antibiofilm strategies.

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来源期刊
Microbiological research
Microbiological research 生物-微生物学
CiteScore
10.90
自引率
6.00%
发文量
249
审稿时长
29 days
期刊介绍: Microbiological Research is devoted to publishing reports on prokaryotic and eukaryotic microorganisms such as yeasts, fungi, bacteria, archaea, and protozoa. Research on interactions between pathogenic microorganisms and their environment or hosts are also covered.
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