Fig. 2.

Ume6 binds the ATG8 promoter and negatively regulates ATG8 transcription. (A) Expression of ATG8p-LacZ in a UME6 deletion strain. Wild-type and ume6Δ cells containing LacZ driven by the ATG8 promoter were grown to midlog phase and switched to nitrogen starvation medium (SD-N) for 2 h. β-galactosidase activity was measured from protein extracts. (B) Protein A–tagged Ume6 binds the ATG8 promoter. ChIP analysis was conducted on two regions of the ATG8 promoter: the URS1 region and a region −3 kb upstream of the ATG8 start codon (-3K), which was used as a negative control. The URS1 region in the INO1 promoter served as a positive control. The ChIP results were normalized to the input DNA and calibrated to the -3K PCR product, which was set to 1.0. Error bars represent the SD of at least three independent experiments.