Manganese transport and virulence in Brucella

布鲁氏菌中锰的转运和毒力

• 批准号: 8847652
• 负责人: ROY M ROOP
• 金额: $ 21.92万
• 依托单位: EAST CAROLINA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-09 至 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8847652
• 关键词: Adverse effects; Affinity; Antibiotics; Award; Bacteria; Bacterial Proteins; Biochemical; Biological Warfare; Bioterrorism; Bradyrhizobium; Brucella; Brucella abortus; Brucella melitensis; Brucellosis; Carbon; Catabolism; Chemistry; Chronic; Defense Mechanisms; Development; Disease; Divalent Cations; Enzymes; Escherichia coli; Exhibits; Exposure to; Glucose; Health; Hexoses; Homologous Protein; Human; Hydrogen Peroxide; In Vitro; Infection; Iron; Knowledge; Maintenance; Mammals; Manganese; Mediating; Metabolic; Metals; Mouse Strains; Mus; Oxidative Stress; Pathogenesis; Pathway interactions; Pentosephosphate Pathway; Phylogenetic Analysis; Physiology; Play; Property; Proteins; Protons; Public Health; Pyruvate Kinase; Regulation; Relative (related person); Research; Role; Stress; Testing; Transcription Coactivator; Vaccines; Virulence; Withdrawal; Work; attenuation; base; biological adaptation to stress; cofactor; deprivation; epimerase; improved; in vitro Assay; in vivo; macrophage; microbial; mutant; oxidative damage; pathogen; prevent; research study; response; ribulose 5-phosphate; symporter; uptake

DESCRIPTION (provided by applicant): The proton symporter MntH serves as the sole high affinity manganese transporter in Brucella strains (2). The extreme attenuation displayed by a B. abortus mntH mutant in experimentally infected mice indicates that Mn plays an exceptionally important role in the virulence of Brucella strains in comparison with other bacterial pathogens that have been examined. Glucose catabolism has recently been shown to be critical for the ability of Brucella strains to persist in alternatively activated macrophages, which in turn is required for the maintenance of chronic infections in experimentally infected mice (51). Brucella strains catabolize glucose exclusively via the pentose phosphate pathway (15), and the activities of two enzymes required for maintaining carbon flow through this pathway, pyruvate kinase (PykM) (20) and ribulose-5-phosphate 3-epimerase (Rpe) (46), are predicted to the particularly susceptible to reduced cellular Mn levels. Consequently, the studies outlined in Specific Aim 1 of this application will test the hypothesis that defective glucose catabolism resulting from reduced PykM and Rpe activities makes a significant contribution to the attenuation exhibited by the B. abortus mntH mutant. Experimental evidence also suggests that Brucella strains have the capacity to increase their MntH-mediated Mn uptake and substitute this metal for iron in cellular proteins as a protective mechanism against oxidative stress, in a similar manner to that recently demonstrated in Escherichia coli (3). The studies described in Specific Aim 2 of this proposal will identify the transcriptional regulator responsible for the H2O2- responsive induction of mntH expression in B. abortus 2308, and determine if this mode of mntH regulation protects this strain from oxidative stress in vitro and is required for its virulence in mice. Results obtained from the proposed studies will begin to explain Mn plays such a critical role in the basic physiology and virulence of Brucella strains. The fact that loss f MntH has such an adverse effect on virulence also makes this transporter an attractive target for the development of improved vaccines and antibiotics to prevent and treat human brucellosis, a major zoonotic disease of worldwide importance.

描述（由申请人提供）：质子同向转运蛋白 MntH 是布鲁氏菌菌株中唯一的高亲和力锰转运蛋白 (2)。流产布鲁氏菌 mntH 突变体在实验感染小鼠中表现出的极度减毒表明，与已检查的其他细菌病原体相比，锰在布鲁氏菌菌株的毒力中发挥着极其重要的作用。最近已证明，葡萄糖分解代谢对于布鲁氏菌菌株在替代激活的巨噬细胞中持续存在的能力至关重要，而巨噬细胞又是实验感染小鼠维持慢性感染所必需的 (51)。布鲁氏菌菌株仅通过磷酸戊糖途径分解代谢葡萄糖 (15)，并且维持通过该途径的碳流所需的两种酶的活性，即丙酮酸激酶 (PykM) (20) 和核酮糖 5-磷酸 3-差向异构酶 (Rpe) (46)，预计特别容易受到细胞 Mn 水平降低的影响。因此，本申请的具体目标 1 中概述的研究将检验以下假设：PykM 和 Rpe 活性降低导致的葡萄糖分解代谢缺陷对流产布鲁氏菌 mntH 突变体表现出的减弱有显着贡献。实验证据还表明，布鲁氏菌菌株有能力增加 MntH 介导的 Mn 摄取，并用这种金属替代细胞蛋白质中的铁，作为对抗氧化应激的保护机制，其方式与最近在大肠杆菌中证明的方式类似 (3)。该提案的具体目标 2 中描述的研究将鉴定负责 B. abortus 2308 中 H2O2 反应性诱导 mntH 表达的转录调节因子，并确定这种 mntH 调节模式是否可以保护该菌株免受体外氧化应激，并且是其在小鼠中的毒力所必需的。从拟议的研究中获得的结果将开始解释锰在布鲁氏菌菌株的基本生理学和毒力中起着如此关键的作用。事实上，f MntH 的缺失对毒力产生如此不利的影响，也使得这种转运蛋白成为开发改进的疫苗和抗生素的有吸引力的目标，以预防和治疗人类布鲁氏菌病，这是一种世界范围内重要的人畜共患疾病。

项目成果

A novel formamidase is required for riboflavin biosynthesis in invasive bacteria.

• DOI: 10.1016/j.jbc.2022.102377
• 发表时间: 2022-09
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Yurgel, Svetlana N.;Johnson, Skylar A.;Rice, Jennifer;Sa, Na;Bailes, Clayton;Baumgartner, John;Pitzer, Josh E.;Roop, R. Martin I. I. I. I.;Roje, Sanja
• 通讯作者: Roje, Sanja

Identifying the region responsible for Brucella abortus MucR higher-order oligomer formation and examining its role in gene regulation.

鉴定负责流产布鲁氏菌 MucR 高阶寡聚体形成的区域并检查其在基因调控中的作用。

• DOI: 10.1038/s41598-018-35432-1
• 发表时间: 2018
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Pirone,Luciano;Pitzer,JoshuaEdison;D'Abrosca,Gianluca;Fattorusso,Roberto;Malgieri,Gaetano;Pedone,EmiliaMaria;Pedone,PaoloVincenzo;Roop2nd,RoyMartin;Baglivo,Ilaria
• 通讯作者: Baglivo,Ilaria

Investigating the roles of the conserved Cu2+-binding residues on Brucella FtrA in producing conformational stability and functionality.

研究布鲁氏菌 FtrA 上保守的 Cu2 结合残基在产生构象稳定性和功能中的作用。

• DOI: 10.1016/j.jinorgbio.2020.111162
• 发表时间: 2020
• 期刊: Journal of inorganic biochemistry
• 影响因子: 3.9
• 作者: Banerjee,Sambuddha;Garrigues,RyanJ;Chanakira,MinaN;Negron-Olivo,JacobJ;Odeh,YasmeneH;Spuches,AnneM;MartinRoop2nd,R;Pitzer,JoshuaEdison;Martin,DanielW;Dasgupta,Saumya
• 通讯作者: Dasgupta,Saumya