Journal of Amino Acids / 2015 / Article / Fig 3

Research Article

Genes of Different Catabolic Pathways Are Coordinately Regulated by Dal81 in Saccharomyces cerevisiae

Figure 3

Role of Dal81 in AGP1 and BAP2 regulation. Wild type cells, expressing the Stp1-HA (DEBY01 strain) fusion protein, were treated or not with leucine. ChIP assays were carried out using antibodies against the HA epitope. qPCR was performed with specific primers (black bars) that amplify a region of AGP1 promoter (a), a region of BAP2 promoter (b), and a region 2.5 kb downstream of UGA4 promoter (white bars) used as a negative control. Results are expressed as the fold change of binding to the promoter of interest and are the mean ± SEM of three independent experiments. mRNA levels of AGP1 (c) and BAP2 (d) were determined in wild type cells (23344c strain), leu3Δ cells (MPY09 strain), and leu3Δ dal81Δ cells (SBCY20 strain). The cells were incubated with or without leucine for 30 minutes. mRNA levels were quantified by RT-qPCR. AGP1 and BAP2 values were normalized with TBP1 and results are the mean ± SEM of three independent experiments.

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