Chemical rescue of mutant proteins in living Saccharomyces cerevisiae cells by naturally occurring small molecules

Intracellular proteins function in a complex milieu wherein small molecules influence protein folding and act as essential cofactors for enzymatic reactions. Thus protein function depends not only on amino acid sequence but also on the concentrations of such molecules, which are subject to wide variation between organisms, metabolic states, and environmental conditions. We previously found evidence that exogenous guanidine reverses the phenotypes of specific budding yeast septin mutants by binding to a WT septin at the former site of an Arg side chain that was lost during fungal evolution. Here we used a combination of targeted and unbiased approaches to look for other cases of “chemical rescue” by naturally occurring small molecules. We report in vivo rescue of hundreds of Saccharomyces cerevisiae mutants representing a variety of genes, including likely examples of Arg or Lys side chain replacement by the guanidinium ion. Failed rescue of targeted mutants highlight features required for rescue, as well as key differences between the in vitro and in vivo environments. Some non-Arg mutants rescued by guanidine likely result from “off-target” effects on specific cellular processes in WT cells. Molecules isosteric to guanidine and known to influence protein folding had a range of effects, from essentially none for urea, to rescue of a few mutants by DMSO. Strikingly, the osmolyte trimethylamine-N-oxide rescued ∼20% of the mutants we tested, likely reflecting combinations of direct and indirect effects on mutant protein function. Our findings illustrate the potential of natural small molecules as therapeutic interventions and drivers of evolution.

Recognition and Status of Response Equipment Related to Chemical Accidents and Terrorism

This study analyzes the perception, status, and problems of domestic firefighters related to chemical accidents and chemical terror response equipment. “How to use on-site response facilities and equipment” was 10% when “0 times” and 28.4% when “1 to 4 times,” indicating that the importance of the ‘experience’ category is greatly increased when there is a lack of relative experience. As for whether measurement equipment is retained, 20.0% of negative responses in the ‘special structure’ were found to differ in function by “Duty.” The effects of equipment-related proficiency and preference appear to be differences in equipment primarily held by special and chemical rescue teams. Problems with on-site response will require advisory experts on post-processing and analyses and the training of professional firefighters. Accordingly, through intensive interest and analyses in the field of special disaster responses, securing expertise, and fostering professional personnel, these problems can be addressed.

Chemical rescue of mutant proteins in living cells by naturally occurring small molecules

Intracellular proteins function in a complex milieu wherein small molecules influence protein folding and act as essential cofactors for enzymatic reactions. Thus protein function depends not only on amino acid sequence but also on the concentrations of such molecules, which are subject to wide variation between organisms, metabolic states, and environmental conditions. We previously found evidence that exogenous guanidine reverses the phenotypes of specific budding yeast septin mutants by binding to a WT septin at the former site of an Arg side chain that was lost during fungal evolution. Here we used a combination of targeted and unbiased approaches to look for other cases of “chemical rescue” by naturally occurring small molecules. We reportin vivorescue of hundreds of yeast mutants representing a variety of genes, including likely examples of Arg or Lys side chain replacement by the guanidinium ion. Failed rescue of targeted mutants highlight features required for rescue, as well as key differences between thein vitroandin vivoenvironments. Some non-Arg mutants rescued by guanidine likely result from “off-target” effects on specific cellular processes in WT cells. Molecules isosteric to guanidine and known to influence protein folding had a range of effects, from essentially none for urea, to rescue of a few mutants by DMSO. Strikingly, the osmolyte trimethylamine-N-oxide rescued ∼20% of the mutants we tested, likely reflecting combinations of direct and indirect effects on mutant protein function. Our findings illustrate the potential of natural small molecules as therapeutic interventions and drivers of evolution.

Analysis of an Event Related to the Uncontrolled Release of Ammonia from a Rail Tanker

Numerical engineering tools are being more and more frequently applied in supporting decisions responsible for rescue operations in terms of tactics and strategy. This elaboration presents one of the applications of such software in the event of uncontrolled releases of hazardous substances. The adoption of numerical modelling as an element which supports leader-ship results in the reduction of operating costs and affects its performance. The example used will show the potential of these solutions to people unrelated directly to chemical and ecological rescue. It may become a basis for developing drills, including evacuation drills, for people responsible for their planning and organization in the railway structures. Keywords: chemical rescue, dispersion modelling, ALOHA