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蓝光对单核细胞增生李斯特菌生长的抑制作用由活性氧介导,并受σB和蓝光传感器Lmo0799的影响。

Blue-Light Inhibition of Listeria monocytogenes Growth Is Mediated by Reactive Oxygen Species and Is Influenced by σB and the Blue-Light Sensor Lmo0799.

作者信息

O'Donoghue Beth, NicAogáin Kerrie, Bennett Claire, Conneely Alan, Tiensuu Teresa, Johansson Jörgen, O'Byrne Conor

机构信息

Bacterial Stress Response Group, Microbiology, School of Natural Sciences, National University of Ireland, Galway, Galway, Ireland.

National Centre for Laser Applications, School of Physics, National University of Ireland, Galway, Galway, Ireland.

出版信息

Appl Environ Microbiol. 2016 Jun 13;82(13):4017-4027. doi: 10.1128/AEM.00685-16. Print 2016 Jul 1.

Abstract

UNLABELLED

Listeria monocytogenes senses blue light via the flavin mononucleotide-containing sensory protein Lmo0799, leading to activation of the general stress response sigma factor SigB (σ(B)). In this study, we investigated the physiological response of this foodborne pathogen to blue light. We show that blue light (460 to 470 nm) doses of 1.5 to 2 mW cm(-2) cause inhibition of growth on agar-based and liquid culture media. The inhibitory effects are dependent on cell density, with reduced effects evident when high cell numbers are present. The addition of 20 mM dimethylthiourea, a scavenger of reactive oxygen species, or catalase to the medium reverses the inhibitory effects of blue light, suggesting that growth inhibition is mediated by the formation of reactive oxygen species. A mutant strain lacking σ(B) (ΔsigB) was found to be less inhibited by blue light than the wild type, likely indicating the energetic cost of deploying the general stress response. When a lethal dose of light (8 mW cm(-2)) was applied to cells, the ΔsigB mutant displayed a marked increase in sensitivity to light compared to the wild type. To investigate the role of the blue-light sensor Lmo0799, mutants were constructed that either had a deletion of the gene (Δlmo0799) or alteration in a conserved cysteine residue at position 56, which is predicted to play a pivotal role in the photocycle of the protein (lmo0799 C56A). Both mutants displayed phenotypes similar to the ΔsigB mutant in the presence of blue light, providing genetic evidence that residue 56 is critical for light sensing in L. monocytogenes Taken together, these results demonstrate that L. monocytogenes is inhibited by blue light in a manner that depends on reactive oxygen species, and they demonstrate clear light-dependent phenotypes associated with σ(B) and the blue-light sensor Lmo0799.

IMPORTANCE

Listeria monocytogenes is a bacterial foodborne pathogen that can cause life-threatening infections in humans. It is known to be able to sense and respond to visible light. In this study, we examine the effects of blue light on the growth and survival of this pathogen. We show that growth can be inhibited at comparatively low doses of blue light, and that at higher doses, L. monocytogenes cells are killed. We present evidence suggesting that blue light inhibits this organism by causing the production of reactive oxygen species, such as hydrogen peroxide. We help clarify the mechanism of light sensing by constructing a "blind" version of the blue-light sensor protein. Finally, we show that activation of the general stress response by light has a negative effect on growth, probably because cellular resources are diverted into protective mechanisms rather than growth.

摘要

未标记

单核细胞增生李斯特菌通过含黄素单核苷酸的传感蛋白Lmo0799感知蓝光,从而导致一般应激反应σ因子SigB(σ(B))的激活。在本研究中,我们调查了这种食源性病原体对蓝光的生理反应。我们发现,1.5至2 mW/cm²的蓝光剂量(460至470 nm)会抑制基于琼脂的培养基和液体培养基上的生长。抑制作用取决于细胞密度,当细胞数量较多时,抑制作用减弱。向培养基中添加20 mM二甲基硫脲(一种活性氧清除剂)或过氧化氢酶可逆转蓝光的抑制作用,这表明生长抑制是由活性氧的形成介导的。发现缺乏σ(B)的突变株(ΔsigB)比野生型受蓝光的抑制作用更小,这可能表明启动一般应激反应存在能量消耗。当向细胞施加致死剂量的光(8 mW/cm²)时,与野生型相比,ΔsigB突变株对光的敏感性显著增加。为了研究蓝光传感器Lmo0799的作用,构建了缺失该基因的突变株(Δlmo0799)或在第56位保守半胱氨酸残基处发生改变的突变株,预计该残基在蛋白质的光循环中起关键作用(lmo0799 C56A)。在蓝光存在下,这两种突变株均表现出与ΔsigB突变株相似的表型,提供了遗传证据表明第56位残基对单核细胞增生李斯特菌的光感知至关重要。综上所述,这些结果表明,单核细胞增生李斯特菌受到蓝光抑制的方式取决于活性氧,并且它们证明了与σ(B)和蓝光传感器Lmo0799相关的明显的光依赖性表型。

重要性

单核细胞增生李斯特菌是一种食源性细菌病原体,可导致人类危及生命的感染。已知它能够感知并对可见光做出反应。在本研究中,我们研究了蓝光对这种病原体生长和存活的影响。我们表明,相对低剂量的蓝光即可抑制生长,而在较高剂量下,单核细胞增生李斯特菌细胞会被杀死。我们提供的证据表明,蓝光通过导致过氧化氢等活性氧的产生来抑制这种生物体。我们通过构建蓝光传感器蛋白的“盲”版本来帮助阐明光感知机制。最后,我们表明光激活一般应激反应对生长有负面影响,可能是因为细胞资源被转移到保护机制而非生长中。

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