The long-term goal of the project is to expand our knowledge regarding the physiology, metabolism and evolutionary adaptation of Methanosarcina species. The primary emphasis of these studies will be the identification and characterization of genes that directly and indirectly effect the utilization of methanol as a growth substrate. These studies are expected to reveal novel aspects of the methylotrophic methanogenic pathway, and lend insight into the integration of methanogenesis into the global metabolism of the organism. Moreover, a clearer understanding of archaeal transcriptional control should result from examination of the highly regulated methylotrophic genes. Lastly, new genetic methods, which may be widely applicable, will be developed during the course of these studies. The proposed experiments will primarily utilize recently developed methods for genetic analysis of Methanosarcina species; however, physiological, biochemical and molecular approaches will complement the genetic approach. This strategy is expected to generate data that are both novel and complementary to the results of preceding studies. These studies will be broadly applicable to the field of methanogenesis, which is important to a number of human-related problems including the production of alternative fuels from biological materials, waste treatment, and global warming. This project will provide training opportunities for two graduate students and several undergraduate students.