Genetic Analysis of Methanogenesis by Methanosarcina species

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The long-term goal of the proposed research is to understand fully the physiology, metabolism and evolutionary adaptation of Methanosarcina species. The primary emphasis of this proposal is the identification and characterization of genes that directly and indirectly affect the use of one-carbon compounds as substrate for growth and methane production. The research will (1) examine how genes are regulated by methanol, (2) examine which genes are regulated by methanol, and (3) develop preliminary data to support the existence of a novel mechanism to regulate production of proteins in Methanosarcina. The proposed experiments will primarily utilize recently developed methods for genetic analysis of Methanosarcina; however, physiological, biochemical and molecular approaches will complement the genetic approach. The intellectual merit of the proposed research is based on the central role of methanogenic microorganisms in a number of human-related problems including the production of alternative fuels from biological materials, agriculture, waste treatment, and global warming. The proposed studies are expected to reveal novel aspects of the methylotrophic pathway by lending insight into the integration of methanogenesis into the global metabolism of the organism. Moreover, a clearer understanding of archaeal transcriptional and translational control should result from examination of the highly regulated methylotrophic genes. The broader impacts of the proposed research include the training of future scientists and academics. Two students will be directly supported by the requested funds. This study is likely to produce new genetic tools and methods that will be useful, and freely available, to the wider microbial research community. Finally, students from other universities may be trained in the methods developed during this research. Facilities, resources and advice will also be provided to local elementary and secondary school teachers who wish to expose their students to the excitement of the microbial world.