CAREER: Iron-sulfur cluster assembly in Bacillus subtilis

职业：枯草芽孢杆菌中的铁硫簇组装

• 批准号: 1750624
• 负责人: Jeffrey Boyd
• 金额: $ 103.37万
• 依托单位: Rutgers University New Brunswick
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1750624&HistoricalAwards=false
• 关键词: CAREER; Iron; sulfur; cluster; assembly

All forms of life require that many of their proteins have non-protein parts known as cofactors added to them so that they can function properly. Many of these cofactors are built from vitamins and minerals that are important in the diet. One such cofactor is the iron-sulfur (Fe-S) cluster, which allows proteins to perform many important types of chemistry. Because free Fe and S are toxic to cells, elaborate protein systems are used to build these cofactors. Once they are built, the Fe-S clusters must be trafficked to, and inserted into, proteins that require them. Cells that cannot properly build Fe-S clusters or maturate Fe-S proteins have chronic complications, which often results in cell death. The nature of this cellular machinery, however, is not well understood, and many questions remain about how organisms build Fe-S clusters and deliver them to proteins that require them for function. Novel Fe-S protein maturation proteins continue to be discovered, highlighting the fact that a global understanding of this process is lacking. The studies to be conducted in this project will impact the understanding of iron-dependent processes from bacteria to humans. The project will also provide unique educational opportunities for two postdoctoral scholars and up to ten undergraduate students. These trainees will be technically and intellectually trained and will receive the necessary guidance and resources to aid their transition into the next phases of their careers. The trainees will focus on enhancing their communication skills, technical skills, teaching skills, and personnel management skills.The Gram-positive bacterium Bacillus subtilis utilizes the SufCDSUB Fe-S cluster biosynthetic system and at least two Fe-S cluster carriers. Additional factors have been identified that have roles in Fe-S protein maturation, including SufT and YlaN; however, the functions of these molecules are unknown. Moreover, the Fe and electron donors for Suf-directed Fe-S cluster synthesis, as well as additional carrier molecules, are unknown. To properly understand how Fe-S proteins are maturated, the components involved must be identified and their functions defined. This project combines classic genetics and biochemistry with state-of-the-art, next-generation technologies and computational biology to 1) determine the physical interactions between SufT, YlaN, and Fe-S cluster synthesis, assembly, and target proteins, 2) quantify the necessity of SufT and alternate Fe-S cluster assembly factors for the function of specific metabolic pathways that require Fe-S enzymes, 3) assign biochemical functions to SufT and YlaN, and 4) model and predict the effect of faulty Fe-S protein maturation and validate the effects in vivo.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

所有形式的生命都需要它们的许多蛋白质具有被称为辅因子的非蛋白质部分，以使它们能够正常工作。这些辅助因子中的许多都是由饮食中重要的维生素和矿物质组成的。其中一个辅因子是铁硫（Fe-S）簇，它允许蛋白质进行许多重要类型的化学反应。由于游离的Fe和S对细胞有毒，因此使用精细的蛋白质系统来构建这些辅因子。一旦它们被构建，Fe-S簇必须被运输到并插入需要它们的蛋白质中。无法正确构建Fe-S簇或成熟Fe-S蛋白的细胞会出现慢性并发症，这通常会导致细胞死亡。然而，这种细胞机制的性质还没有得到很好的理解，关于生物体如何构建Fe-S簇并将它们传递给需要它们发挥功能的蛋白质，仍然存在许多问题。新的Fe-S蛋白成熟蛋白不断被发现，这突出了一个事实，即缺乏对这一过程的全面了解。在这个项目中进行的研究将影响对从细菌到人类的铁依赖过程的理解。该项目还将为两名博士后学者和多达10名本科生提供独特的教育机会。这些受训人员将接受技术和智力培训，并将获得必要的指导和资源，以帮助他们过渡到职业生涯的下一个阶段。培训人员将重点提高他们的沟通能力、技术能力、教学能力和人事管理能力。革兰氏阳性菌枯草芽孢杆菌利用SufCD 4 Fe-S簇生物合成系统和至少两种Fe-S簇载体。已经确定了在Fe-S蛋白成熟中起作用的其他因子，包括SufT和YlaN;然而，这些分子的功能尚不清楚。此外，用于Suf定向Fe-S簇合物合成的Fe和电子供体以及另外的载体分子是未知的。为了正确理解Fe-S蛋白是如何成熟的，必须鉴定所涉及的组分并定义其功能。该项目将经典遗传学和生物化学与最先进的新一代技术和计算生物学相结合，以1）确定SufT，YlaN和Fe-S簇合成，组装和靶蛋白之间的物理相互作用，2）量化SufT和替代Fe-S簇组装因子对于需要Fe-S酶的特定代谢途径的功能的必要性，3）为SufT和YlaN分配生化功能，4）模拟和预测错误的Fe-S蛋白成熟的影响，并在体内验证这些影响。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

Superoxide Dismutase and Pseudocatalase Increase Tolerance to Hg(II) in Thermus thermophilus HB27 by Maintaining the Reduced Bacillithiol Pool

• DOI: 10.1128/mbio.00183-19
• 发表时间: 2019-03-01
• 期刊: MBIO
• 影响因子: 6.4
• 作者: Norambuen, Javiera;Hanson, Thomas E.;Boyd, Jeffrey M.
• 通讯作者: Boyd, Jeffrey M.

Contribution of YjbIH to Virulence Factor Expression and Host Colonization in Staphylococcus aureus

• DOI: 10.1128/iai.00155-19
• 发表时间: 2019-06-01
• 期刊: INFECTION AND IMMUNITY
• 影响因子: 3.1
• 作者: Austin, Crystal M.;Garabaglu, Siamak;Bose, Jeffrey L.
• 通讯作者: Bose, Jeffrey L.

Genetic approaches to uncover gene products involved in iron-sulfur protein maturation.

揭示与铁硫蛋白成熟有关的基因产物的遗传学方法。

• DOI: 10.1007/978-1-0716-1605-5_3
• 发表时间: 2021
• 期刊: Methods in molecular biology
• 作者: Valerie Carabetta, Karla Esquilin-Lebron