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[具体物质]在空肠弯曲菌蛋氨酸代谢及最佳鸡定殖中的作用

Role of in Methionine Metabolism and Optimal Chicken Colonization in Campylobacter jejuni.

作者信息

Ruddell Brandon, Hassall Alan, Sahin Orhan, Zhang Qijing, Plummer Paul J, Kreuder Amanda J

机构信息

Department of Veterinary Microbiology and Preventive Medicine, College of Veterinary Medicine, Iowa State University, Ames, Iowa, USA.

Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, Iowa, USA.

出版信息

Infect Immun. 2020 Dec 15;89(1). doi: 10.1128/IAI.00542-20.

Abstract

is a zoonotic pathogen and is one of the leading causes of human gastroenteritis worldwide. IA3902 (representative of the sheep abortion clone) is genetically similar to W7 (representative of strain type NCTC 11168); however, there are significant differences in the ability of mutants of these strains to colonize chickens. LuxS is essential for the activated methyl cycle and generates homocysteine for conversion to l-methionine. Comparative genomics identified differential distribution of the genes and , which function to convert homoserine for downstream production of l-methionine, between IA3902 and W7, which could enable a secondary pathway for l-methionine biosynthesis in a W7 Δ but not in an IA3902 Δ strain. To test the hypothesis that the genes and contribute to l-methionine production and chicken colonization by , we constructed two mutants for phenotypic comparison, the W7 Δ Δ and IA3902 Δ:: mutants. Quantitative reverse transcription-PCR and tandem mass spectrometry protein analysis were used to validate MetAB transcription and translation as present in the IA3902 Δ:: mutant and absent in the W7 Δ Δ mutant. Time-resolved fluorescence resonance energy transfer fluorescence assays demonstrated that l-methionine and -adenosyl methionine concentrations decreased in the W7 Δ Δ mutant and increased in the IA3902 Δ:: mutant. Assessment of chicken colonization revealed that the IA3902 Δ:: strain partially rescued the colonization defect of the IA3902 Δ strain, while the W7 Δ Δ strain showed significantly decreased colonization compared to that of the wild-type and the W7 Δ strain. These results indicate that the ability to maintain l-methionine production , conferred by and in the absence of , is critical for normal chicken colonization by .

摘要

是一种人畜共患病原体,是全球人类肠胃炎的主要病因之一。IA3902(绵羊流产克隆株的代表)在基因上与W7(NCTC 11168菌株类型的代表)相似;然而,这些菌株的突变体在定殖于鸡体内的能力方面存在显著差异。LuxS对活化甲基循环至关重要,并产生同型半胱氨酸以转化为L-蛋氨酸。比较基因组学确定了IA3902和W7之间在将高丝氨酸转化为L-蛋氨酸下游产物的基因和功能上的差异分布,这可能为W7 Δ菌株而非IA3902 Δ菌株中的L-蛋氨酸生物合成提供一条次要途径。为了验证基因和有助于产生L-蛋氨酸并通过定殖于鸡体内的假设,我们构建了两个突变体用于表型比较,即W7 ΔΔ和IA3902 Δ::突变体。定量逆转录PCR和串联质谱蛋白质分析用于验证IA3902 Δ::突变体中存在而W7 ΔΔ突变体中不存在的MetAB转录和翻译。时间分辨荧光共振能量转移荧光测定表明,W7 ΔΔ突变体中L-蛋氨酸和S-腺苷甲硫氨酸浓度降低,而IA3902 Δ::突变体中浓度升高。对鸡定殖的评估表明,IA3902 Δ::菌株部分挽救了IA3902 Δ菌株的定殖缺陷,而W7 ΔΔ菌株与野生型和W7 Δ菌株相比,定殖显著降低。这些结果表明,在不存在时由和赋予的维持L-蛋氨酸产生的能力对于正常定殖于鸡体内至关重要。

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