Exploiting Violet-Blue Light to Kill Campylobacter jejuni: Analysis of Global Responses, Modeling of Transcription Factor Activities, and Identification of Protein Targets.
Exploiting Violet-Blue Light to Kill Campylobacter jejuni: Analysis of Global Responses, Modeling of Transcription Factor Activities, and Identification of Protein Targets.
复制标题
DOI：
10.1128/msystems.00454-22
复制
发表时间：
2022-08-30
期刊：
影响因子：
6.4
通讯作者：
中科院分区：
文献类型：
--
作者：
空肠弯曲菌是一种微需氧食源性人畜共患病病原体，是引起细菌性胃肠炎的主要原因。许多菌株对抗生素的耐药性越来越强，需要新的控制方法来减少食物链污染。一种可能性是使用紫蓝（VB）光的光动力灭活（PDI），C。空肠高度敏感。在这里，我们表明，黄素和原卟啉IX是主要的内源性光敏剂和细胞暴露于VB光增加细胞内活性氧（ROS）的高水平，所示的二氯二氢荧光素报告。对于一种呼吸氧气的细菌来说，C。空肠采用了几个ROS敏感的铁硫簇酶在中央代谢途径，我们表明，VB光导致丙酮酸和2-酮戊二酸氧化还原酶快速失活，从而中断柠檬酸循环。暴露于VB光的细胞也会从C型细胞色素中失去血红素，限制电子传递，这可能是由于血红素连接半胱氨酸残基的不可逆氧化。通过RNAseq和概率建模对全局基因表达变化的评估显示了对VB光的两阶段蛋白质损伤/氧化应激反应，由特定调节剂驱动，包括HspR，PerR，Fur和RacR。缺失突变体分析表明，超氧化物歧化酶和细胞色素CccA是特别重要的VB光的生存和废除阻遏的伴侣和抗氧化应激基因的HcrA，HspR，或PerR增加耐受VB光。我们的结果解释了C.空肠到VB轻，并提供新的见解，可能有助于利用PDI的新的食物链干预措施，以控制这种病原体。重要性弯曲杆菌病由C.空肠是全世界最普遍的人畜共患肠道疾病之一，给人类健康和经济造成巨大负担，抗药性菌株的出现加剧了这一负担。迫切需要新的干预措施来减少食物链污染。虽然众所周知紫外线具有杀菌作用，但它具有高度致突变性，并且在食品生产设施中连续暴露会产生问题;相比之下，窄光谱紫蓝（VB）光更安全。我们证实C.空肠对VB光高度敏感，然后确定了一些参与响应光氧化损伤的全球调节网络。受损细胞成分的识别支持了开发基于VB光的技术的商业应用的努力。
Campylobacter jejuni is a microaerophilic foodborne zoonotic pathogen of worldwide concern as the leading cause of bacterial gastroenteritis. Many strains are increasingly antibiotic resistant and new methods of control are required to reduce food-chain contamination. One possibility is photodynamic inactivation (PDI) using violet-blue (VB) light, to which C. jejuni is highly susceptible. Here, we show that flavin and protoporphyrin IX are major endogenous photosensitizers and that exposure of cells to VB light increases intracellular reactive oxygen species (ROS) to high levels, as indicated by a dichlorodihydrofluorescein reporter. Unusually for an oxygen-respiring bacterium, C. jejuni employs several ROS-sensitive iron-sulfur cluster enzymes in central metabolic pathways; we show that VB light causes rapid inactivation of both pyruvate and 2-oxoglutarate oxidoreductases, thus interrupting the citric acid cycle. Cells exposed to VB light also lose heme from c-type cytochromes, restricting electron transport, likely due to irreversible oxidation of heme-ligating cysteine residues. Evaluation of global gene expression changes by RNAseq and probabilistic modeling showed a two-stage protein damage/oxidative stress response to VB light, driven by specific regulators, including HspR, PerR, Fur, and RacR. Deletion mutant analysis showed that superoxide dismutase and the cytochrome CccA were particularly important for VB light survival and that abolishing repression of chaperones and oxidative stress resistance genes by HcrA, HspR, or PerR increased tolerance to VB light. Our results explain the high innate sensitivity of C. jejuni to VB light and provide new insights that may be helpful in exploiting PDI for novel food-chain interventions to control this pathogen. IMPORTANCE Campylobacteriosis caused by C. jejuni is one of the most widespread zoonotic enteric diseases worldwide and represents an enormous human health and economic burden, compounded by the emergence of antibiotic-resistant strains. New interventions are urgently needed to reduce food-chain contamination. Although UV light is well known to be bactericidal, it is highly mutagenic and problematic for continuous exposure in food production facilities; in contrast, narrow spectrum violet-blue (VB) light is much safer. We confirmed that C. jejuni is highly susceptible to VB light and then identified some of the global regulatory networks involved in responding to photo-oxidative damage. The identification of damaged cellular components underpins efforts to develop commercial applications of VB light-based technologies.
登录
查看更多内容
DOI：
10.1093/bioinformatics/btu638
发表时间：
2015-01-15
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Anders S;Pyl PT;Huber W
通讯作者：
Huber W
影响因子：
3.3
作者：
Dai, Tianhong;Huang, Ying-Ying;Hamblin, Michael R.
通讯作者：
Hamblin, Michael R.
影响因子：
4.3
作者：
Fyrestam J;Bjurshammar N;Paulsson E;Johannsen A;Östman C
通讯作者：
Östman C
影响因子：
--
作者：
Adair TL;Drum BE
通讯作者：
Drum BE
影响因子：
5.2
作者：
Gundogdu O;da Silva DT;Mohammad B;Elmi A;Mills DC;Wren BW;Dorrell N
通讯作者：
Dorrell N