Project description:Iron is an essential cofactor for enzymes involved in numerous cellular processes. We analyzed the metabolomes and transcriptomes of yeast grown in iron-rich and iron-poor media to determine which biosynthetic processes are altered when iron availability falls. Saccharomyces cerevisiae DBY7286 strain was grown from very low density to mid-log phase (A600 = 0.5, approximately 18 hrs.) in defined-iron SD minimal medium containing only the supplements necessary to meet auxotrophic requirements. Defined-iron SD minimal media were prepared with yeast nitrogen base lacking iron and copper, supplemented with 1 µM copper sulfate, 25 mM MES pH 6.1, 1 mM Ferrozine (Fluka), and the indicated concentrations of ferrous ammonium sulfate 10 µM (low iron) or 300 µM (high iron). All cells were grown at 30°C with shaking and four independent cultures were prepared for each growth condition
Project description:Dynamic transcriptional response of S. cerevisiae cells to copper impulse was investigated in both HO deletion strain used as the reference strain and the mutant strain lacking CCC2 gene which were grown in continuous cultures using a copper-deficient defined medium. Copper was introduced into the medium as an impulse so as to reach a copper sulfate concentration of 0.5 mM. Samples were collected within the first two hours following the copper addition (1st, 5th, 10th, 15th, 20th, 25th, 30th, 60th, 120th minutes), in addition to the steady-state sampling.
