2008 — 2009 |
Cooper, Sara J |
F32Activity Code Description: To provide postdoctoral research training to individuals to broaden their scientific background and extend their potential for research in specified health-related areas. |
Assessing Effects of Genetics and Environment On Metabolism in S Cerevisiae @ University of Washington
[unreadable] DESCRIPTION (provided by applicant): Metabolism encompasses all the processes by which a cell generates energy and other essential molecules from nutrients. These pathways rely on hundreds of genes and involve thousands of small molecule intermediates, vitamins and cofactors. Small-scale analysis of metabolites is already a common practice for clinical diagnosis. Increased interest in small molecules has led to development of technologies that allow high-throughput profiling of metabolic intermediates. These methods have already led to new diagnostic techniques for kidney cancer. To further our understanding of the roles for small molecule metabolites, I have begun using capillary electrophoresis to profile metabolites in the yeast Saccharomyces cerevisiae. Yeast share all major metabolic pathways with humans and provide the advantages of a well-studied model organism. Based on the similarities, results in yeast can be easily translated to human biology. Preliminary studies demonstrate that this profiling method is simple, quantitative and easily amenable to automation. First, I will identify derivatization reagents that allow me to observe a wide variety of metabolites. Second, I will characterize yeast genes that affect the levels of cellular metabolites relevant to essential metabolic pathways. Third, I will profile metabolites during response to environmental stimuli and during cellular processes such as the cell cycle. Fourth, I will use yeast to determine how polymorphisms in human genes affect these pathways in human metabolism. These data will provide a clearer picture of the metabolic network in yeast and human, yielding insights into the biology of metabolic processes in human health and disease. PUBLIC HEALTH RELEVANCE: The basic processes of metabolism lie beneath every process of cell biology. By examining and understanding a cell's metabolic response to stimuli, we will gain insights into diverse biological processes. In addition, comparing normal cellular changes with the changes that occur during cells affected by cancer or diabetes provides a potentially novel mechanism for disease prevention and treatment. [unreadable] [unreadable] [unreadable]
|
0.954 |
2010 |
Cooper, Sara J |
P41Activity Code Description: Undocumented code - click on the grant title for more information. |
Seminars Given by Sara Cooper @ University of Washington
This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. HudsonAlpha Institute Huntsville, AL 10/12/09 Medical College of Wisconsin Milwaukee, WI 6/29/09 Metabolomics Meeting Edmonton, Alberta Canada 8/30/09 Vanderbilt University Nashville, TN 10/30/09 Washington University St. Louis, MO 12/3/09 University of Michigan Ann Arbor, MI 1/11/10
|
0.954 |
2010 |
Cooper, Sara J |
P41Activity Code Description: Undocumented code - click on the grant title for more information. |
Small Molecule Modulators of Statin Response in Yeast @ University of Washington
This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We established a yeast-based screen to explore how endogenous metabolites interact with lovastatin, a cholesterol-lowering drug. Lovastatin and other statins inhibit HMG-CoA reductase, which carries out one of the first steps in the sterol biosynthesis pathway. In the yeast Saccharomyces cerevisiae, treatment with a statin results in reduced growth. By treating yeast with lovastatin in combination with a small library of diverse metabolites, we found that copper and zinc ions reduced the ability of the statin to inhibit growth. We analyzed both gene expression and levels of ergosterol and its precursor compounds after treatment with both lovastatin and either of these metals. Statin treatment alone induced sterol and cell wall biosynthesis genes and decreased the levels of ergosterol and its precursors. Metal treatment alone induced many metal homeostasis genes and upregulated a subset of sterol biosynthesis genes, resulting in significantly increased sterol levels. The combination of the drug plus metal synergistically upregulated some sterol biosynthesis genes, resulting in greater flux through the sterol biosynthesis pathway and a concomitant increase in ergosterol levels. The cell responds to the ergosterol deprivation induced by a statin with an increase in ergosterol biosynthesis, and copper and zinc can enhance this response. In cultured mammalian cells, these two metals also rescued statin growth inhibition. Thus, copper and zinc levels could have relevance to statin response in humans. Metabolite perturbation screens such as the one presented here can be performed against nearly any yeast phenotype, and therefore are applicable to a wide range of problems.
|
0.954 |
2010 |
Cooper, Sara J |
P41Activity Code Description: Undocumented code - click on the grant title for more information. |
Yeast Metabolomics by Two-Dimensional Gas Chromatography and Mass Spectrometry @ University of Washington
This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Metabolomics is the unbiased study of small molecules in biological samples. We have previously generated methods for targeted metabolite profiling of amino acids and other classes of small molecules in yeast. Now we have focused our efforts on using two-dimensional gas chromatography with mass spectrometry for an untargeted metabolomics approach in yeast. We have optimized these protocols for the quantification of approximately 150-200 different small molecule metabolites found in yeast. We are applying these methods for the characterization of natural variation in wild yeast strains. In collaboration with other members of the yeast resource center, we are measuring metabolic profiles for 38 individual strains grown under chemostat controlled conditions. Our data in combination with gene expression and proteomics data could provide a more complete network of gene regulation, gene expression and cellular metabolism differ among these strains. This project is just one example of the applications for this technology which can easily be applied to human biofluids and tissue culture samples.
|
0.954 |