Real Time Analysis Of Gene Expression In Living Yeast Cells: Novel Approach For Accurate Cell Damage Detection | 17135
Journal of Biotechnology & Biomaterials
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Microorganisms respond to environmental stress by the activation of gene expression programs in order to adapt and
survive. Fine tuned transcriptional activation in response to stress is the result of dynamic interactions of transcription
factors with specific promoter binding sites. A time resolved luciferase reporter assay in living yeast cells to gain insights
into how osmotic, oxidative and nutritional stress signals modulate gene expression in a dose sensitive manner was used.
Specifically, the dose response behavior of four different natural promoters (
GRE2, CTT1, SOD2
) reveals differences
in their sensitivity and dynamics to salt and oxidative stimuli. Characteristic dose response profiles are also obtained for
artificial promoters driven by only one type of stress regulated consensus element, such as CRE, STRE or AP-1 sites. It has
been shown that the stress tolerance of the cell critically modulates the dynamics of its transcriptional response in the case
of oxidative stress. Moreover, it was identified different regulatory elements in the promoters of oxidative and osmotic stress
response genes that are highly specific for the cell damage. In fact we use this technology to distinguish the cause of oxidative
harm of different molecules such as toxins or oxidative compounds, in order to understand the biological target of injury,
and therefore infer the molecular mechanisms of cellular defense. Real time analysis of gene expression in living yeast cells is
extremely sensitive and requires low concentrations of the chemical species studied, thus this technology allows the study of
cell damage at physiological levels.
Amparo Pascual-Ahuir Gineris associate professor at the Polytechnic University of Valencia (UPV) department of Biochemistry and Molecular Biology, and
researcher at the Institute of Plant Molecular and Cellular Biology (IBMCP), department of abiotic stress. She has completed his PhD at the UPV in 2001 in the field
of transcriptional regulation of genes under stress response. She spent three years as post-doc at Harvard University, Department of Genetics, granted by a Human
Frontiers Science Program Long term Fellowship. She is Faculty member of the UPV since 2007, and in collaboration with Dr. Proft, she is leading a research group
interested in the study of molecular circuits involved in stress response.
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