EF-hand calcium sensor, EfhP, controls transcriptional regulation of iron uptake by calcium in Pseudomonas aeruginosa.

IF 5.1 1区 生物学 Q1 MICROBIOLOGY mBio Pub Date : 2024-11-13 Epub Date: 2024-10-22 DOI:10.1128/mbio.02447-24
Jacob Burch-Konda, Biraj B Kayastha, Myriam Achour, Aya Kubo, Mackenzie Hull, Reygan Braga, Lorelei Winton, Rendi R Rogers, Erika I Lutter, Marianna A Patrauchan
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Abstract

The human pathogen Pseudomonas aeruginosa (Pa) poses a major risk for a range of severe infections, particularly lung infections in patients suffering from cystic fibrosis (CF). As previously reported, the virulent behavior of this pathogen is enhanced by elevated levels of Ca2+ that are commonly present in CF nasal and lung fluids. In addition, a Ca2+-binding EF-hand protein, EfhP (PA4107), was partially characterized and shown to be critical for the Ca2+-regulated virulence in P. aeruginosa. Here, we describe the rapid (10 min, 60 min), and adaptive (12 h) transcriptional responses of PAO1 to elevated Ca2+ detected by genome-wide RNA sequencing and show that efhP deletion significantly hindered both rapid and adaptive Ca2+ regulation. The most differentially regulated genes included multiple Fe sequestering mechanisms, a large number of extracytoplasmic function sigma factors (ECFσ), and several virulence factors, such as the production of pyocins. The Ca2+ regulation of Fe uptake was also observed in CF clinical isolates and appeared to involve the global regulator Fur. In addition, we showed that the efhP transcription is controlled by Ca2+ and Fe, and this regulation required a Ca2+-dependent two-component regulatory system CarSR. Furthermore, the efhP expression is significantly increased in CF clinical isolates and upon pathogen internalization into epithelial cells. Overall, the results established for the first time that Ca2+ controls Fe sequestering mechanisms in P. aeruginosa and that EfhP plays a key role in the regulatory interconnectedness between Ca2+ and Fe signaling pathways, the two distinct and important signaling pathways that guide the pathogen's adaptation to the host.IMPORTANCEPseudomonas aeruginosa (Pa) poses a major risk for severe infections, particularly in patients suffering from cystic fibrosis (CF). For the first time, kinetic RNA sequencing analysis identified Pa rapid and adaptive transcriptional responses to Ca2+ levels consistent with those present in CF respiratory fluids. The most highly upregulated processes include iron sequestering, iron starvation sigma factors, and self-lysis factors pyocins. An EF-hand Ca2+ sensor, EfhP, is required for at least 1/3 of the Ca2+ response, including the majority of the iron uptake mechanisms and the production of pyocins. Transcription of efhP itself is regulated by Ca2+ and Fe, and increases during interactions with host epithelial cells, suggesting the protein's important role in Pa infections. The findings establish the regulatory interconnectedness between Ca2+ and iron signaling pathways that shape Pa transcriptional responses. Therefore, understanding Pa's transcriptional response to Ca2+ and associated regulatory mechanisms will serve in the development of future therapeutics targeting Pa's dangerous infections.

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EF 手钙传感器 EfhP 控制铜绿假单胞菌中钙对铁吸收的转录调控。
人类病原体铜绿假单胞菌(Pa)对一系列严重感染,尤其是囊性纤维化(CF)患者的肺部感染构成重大风险。据先前报道,CF 鼻腔和肺液中常见的 Ca2+ 水平升高会增强这种病原体的毒性。此外,一种钙2+结合EF-手蛋白EfhP(PA4107)已被部分鉴定并证明对铜绿假单胞菌的钙2+调控毒力至关重要。在这里,我们描述了通过全基因组 RNA 测序检测到的 PAO1 对 Ca2+ 升高的快速(10 分钟、60 分钟)和适应性(12 小时)转录反应,并表明删除 EfhP 会显著阻碍快速和适应性 Ca2+ 调节。调节差异最大的基因包括多种铁螯合机制、大量胞质外功能σ因子(ECFσ)和几种毒力因子,如产生焦蛋白。在CF临床分离株中也观察到了Ca2+对铁吸收的调控,而且似乎涉及全局调控因子Fur。此外,我们还发现efhP的转录受Ca2+和Fe的控制,而这种调控需要一个依赖于Ca2+的双组分调控系统CarSR。此外,efhP的表达在CF临床分离株中和病原体内化到上皮细胞时显著增加。总之,研究结果首次证实了 Ca2+ 控制铜绿假单胞菌的铁螯合机制,而 EfhP 在 Ca2+ 和铁信号通路之间的相互联系中发挥着关键作用,这两种不同的重要信号通路指导着病原体对宿主的适应。动力学 RNA 测序分析首次发现了铜绿假单胞菌对 Ca2+ 水平的快速适应性转录反应,这种反应与 CF 呼吸液中的 Ca2+ 水平一致。上调幅度最大的过程包括铁螯合、铁饥饿sigma因子和自裂解因子pyocins。至少有 1/3 的 Ca2+ 反应需要 EF 手 Ca2+ 传感器 EfhP,包括大多数铁吸收机制和焦蛋白的产生。efhP本身的转录受Ca2+和铁的调控,并在与宿主上皮细胞相互作用时增加,这表明该蛋白在Pa感染中发挥着重要作用。这些发现确定了影响 Pa 转录反应的 Ca2+ 和铁信号通路之间的相互调控关系。因此,了解 Pa 对 Ca2+ 的转录反应及相关调控机制将有助于开发未来针对 Pa 危险感染的治疗药物。
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来源期刊
mBio
mBio MICROBIOLOGY-
CiteScore
10.50
自引率
3.10%
发文量
762
审稿时长
1 months
期刊介绍: mBio® is ASM''s first broad-scope, online-only, open access journal. mBio offers streamlined review and publication of the best research in microbiology and allied fields.
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