Developing Methanosarcina spp. as a model system to study cytochromes c and their role in archaeal methane metabolism

项目总结/摘要 细胞色素c在甲烷代谢古菌中对甲烷的产生和消耗至关重要 再加上增长和节能。总的来说，甲烷代谢古菌介导了甲烷的净通量。 甲烷释放到大气中，从而对全球甲烷循环和气候产生重大影响 变化基于基因组研究，细胞色素c已被假设在以下方面发挥重要作用： 然而，甲烷代谢的基本分子机制仍然难以捉摸，主要是由于 缺乏完善的遗传模型系统。即使是尝试用一种新的方法来研究古细菌细胞色素c， 替代方法例如在良好建立的细菌系统中的异源表达尚未被证实 成功到目前为止。拟议研究的主要重点是开发一种遗传上易于处理的产甲烷菌 古细菌，Methanosarcina acetivorans，作为功能表征古细菌细胞色素c和 获得生理学的见解细胞色素c在甲烷代谢中的作用，在不同的古细菌 物种 细胞色素c是普遍存在的电子转移蛋白，其需要共价连接至其血红素辅因子， 这个过程叫做细胞色素C生物合成。细胞色素c生物合成途径与 感兴趣的细胞色素c对于在异源细胞中成功产生功能性细胞色素c是关键的。 主持人利用遗传和生物化学工具，我最近的特点细胞色素c生物合成途径 产甲烷古菌模式菌醋酸甲烷八叠球菌利用这些知识，该项目旨在 发展M.乙酸食腐菌作为产生和功能表征古细菌细胞色素c的遗传底盘 来自不同的古细菌物种。该项目的目标1将在功能上描述关键的 细胞色素c属于产甲烷古菌或产甲烷菌，Aim2将研究这些蛋白质 使用体外和体内分析从消耗甲烷的古细菌中分离出的。这项研究将提高我们的 了解甲烷代谢，并导致古细菌宿主的发展，以研究细胞色素c 古细菌的蛋白质最终，从这些研究中获得的知识可以用于开发 我们的目标是为全球气候危机提供可持续的解决方案，并减轻其对人类健康的有害影响。

Project Summary/Abstract Cytochromes c are crucial in methane-metabolizing archaea for the production and consumption of methane coupled to growth and energy conservation. Overall, methane-metabolizing archaea mediate the net flux of methane released into the atmosphere and thus, significantly impact the global methane cycle and climate change. Based on genomic studies, Cytochromes c have been hypothesized to play an important role in methane metabolism however the underlying molecular mechanisms remain elusive, primarily due to the absence of a well-developed genetic model system. Even attempts to study archaeal cytochrome c using an alternative approach such as heterologous expression in well-established bacterial systems have not proven successful so far. The main focus of the proposed research is to develop a genetically tractable methanogenic archaeon, Methanosarcina acetivorans, as a platform to functionally characterize archaeal cytochrome c and gain physiological insights into the role of cytochrome c in methane metabolism across different archaeal species. Cytochromes c are ubiquitous electron transfer proteins that require a covalent attachment to its heme co-factor, a process called cytochrome c biogenesis. Coordination between the cytochrome c biogenesis pathway and the cytochrome c of interest is critical for the successful production of a functional cytochrome c in a heterologous host. Using genetic and biochemical tools, I have recently characterized the cytochrome c biogenesis pathway in the model methanogenic archaeon, Methanosarcina acetivorans. Using this knowledge, the project aims to develop M. acetivorans as a genetic chassis to produce and functionally characterize archaeal cytochrome c from diverse archaeal species. Aim1 of the project will functionally characterize the crucial cytochromes c belonging to methane-producing archaea or methanogens, and Aim2 will study these proteins from methane-consuming archaea using both in vitro and in vivo analyses. This research will improve our understanding of methane metabolism and lead to the development of an archaeal host to study cytochrome c proteins from archaea. Ultimately, the knowledge garnered from these studies can be used to develop sustainable solutions for the global climate crisis and mitigate its harmful impacts on human health.