DNA Uptake By Naturally Competent Archaea

微生物可以通过一种称为转化的过程直接从其环境中获得遗传物质。这些生物属于两大类群，细菌和古生菌。细菌的转化得到了很好的研究，但关于古生菌是如何发挥作用的知之甚少，尽管古生菌代表了地球上至少三分之一的生物多样性，并在大型工业和市政生物反应器中催化了全球生物地球化学循环中的基本反应。为了解决这一不足，这一提议将是古生菌成员转化的第一个特征。通过这项工作，将识别进行这一过程所需的必要基因和蛋白质。这些结果可能会进一步努力通过基因技术来研究古细菌中的生物。这项研究有可能直接和积极地影响古细菌催化的任何过程的研究。此外，转化在自然种群中很重要，这里产生的结果将有助于更广泛地了解环境中的基因流动。最后，作为这项研究的一部分开发的技术将通过研究生培训和加强本科生和研究生课堂上的动手实验，直接促进下一代科学家的教育。该项目将描述古生物领域中自然转化的过程--有能力的生物体直接从环境中摄取DNA并将其纳入基因组。这将通过识别和表征古生菌DNA摄取机制的组件来实现，该机制涉及三个不同的转化步骤：DNA的细胞外结合、跨细胞膜的运输和细胞质中DNA的维持。识别这些步骤中涉及的基因和蛋白质将有助于对有机体吸收环境DNA的能力进行电子预测。该项目将利用转座子突变、体外生化分析和表型分析来表征maripaludis甲烷球菌的转化，这是一种研究古细菌领域成员的模式生物。为了进一步促进古生物转化机制的表征，本提案中的工作将开发和利用厌氧古菌的活细胞荧光成像系统来分析合格考古中的蛋白质定位和蛋白质-蛋白质相互作用。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

Microorganisms can acquire genetic material directly from their environment through a process called transformation. These organisms belong to two broad groups, the bacteria and the archaea. Transformation is well studied in bacteria, but little is known about how this works in archaea, despite the fact that archaea represent at least a third of the biodiversity on Earth and catalyze essential reactions in both global biogeochemical cycles in large-scale industrial and municipal bioreactors. To address this deficiency, this proposal will be the first characterization of transformation in a member of the archaea. Through this work, the necessary genes and proteins needed to carry out this process will be identified. These results may further efforts to study organisms from the archaea through genetic techniques. This research has the potential to impact directly and positively research in any process that archaea catalyze. Additionally, transformation is important in natural populations, and the results generated here will help develop a broader understanding of gene flow in the environment. Finally, the techniques developed as part of this research will directly contribute to the education of the next generation of scientists through graduate student training and enhancing hands-on experimentation in undergraduate and graduate classrooms.This project will characterize the process of natural transformation - the direct uptake and genomic incorporation of DNA from the environment by competent organisms - in the archaeal domain. This will be accomplished by identifying and characterizing components of the archaeal DNA uptake machinery involved in three distinct steps of transformation: extracellular binding of DNA, transport across the cell membrane, and maintenance of DNA in the cell cytoplasm. Identifying the genes and proteins involved in these steps will facilitate in silico predictions of an organism’s ability to take up environmental DNA. This project will leverage transposon mutagenesis, in vitro biochemical analysis, and phenotypic assays to characterize transformation in Methanococcus maripaludis, a model organism for the study of members of the archaeal domain. To further facilitate the characterization of archaeal transformation machinery, the work in this proposal will develop and utilize a system for live cell fluorescent imaging of anaerobic archaea to analyze protein localization and protein-protein interactions in competent archaea.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.