This is the final technical report for a project involving the study of stress response systems in the radiation-resistant bacterium, Deinococcus radiodurans. Three stresses of importance for a mixed waste treatment strain were studied, heat shock, solvent shock, and phosphate starvation. In each case, specific genes involved in the ability to survive the stress were identified using a systems biology approach, and analysis of mutants was used to understand mechanisms. This study has led to increased understanding of the ways in which a potential treatment strain could be manipulated to survive multiple stresses for treatment of mixed wastes.

In the future, environmental concerns will mandate that manufacturing processes shift towards the use of renewable resources and the minimization of wastes, especially hazardous wastes. One-carbon compounds are of interest as feedstocks for synthesis of chemicals and materials, because they represent a relatively inexpensive, abundant and renewable resource. In addition, the environmentally-benign characteristics of microbial processes make them of interest as part of a long-term waste-minimization strategy for industry. The concept that methylotrophic bacteria could serve as non-polluting multistage catalysts to generate chemicals and materials using C1 compounds as feedstocks is a highly attractive one. In order to develop production strains of methylotrophs, it is necessary to understand and manipulate central methylotrophic pathways. One of the most important of these is the methanol oxidation, or Mox system. In this project, we are studying the promoters and transcriptional regulation of this 25-gene system in Methylobacterium extorquens AM1, a facultative methanol-utilizer. We have addressed the significance of a hexanucleotide sequence upstream of all mox promoters and have shown that it is required for activity of these promoters using both deletion and mutational analyses. In addition, we have identified a putative hairpin structure in the RNA leader region of the mxa promoter that …

This report discusses issues related to peace in the Middle East, including post-Gulf War negotiations between Israel and the Palestinians, Jordan, Syria, and Lebanon; U.S. interest in these and continuing negotiations and peacetalks between Middle Eastern countries; and the role of the U.S., the U.N., the European Union, and Russia in Middle Eastern relations.