Figure 4.—

Dal81 is required for the efficient promoter binding of latent and processed forms of Stp1. ChIP analysis of Stp1 association with AGP1 and GNP1 promoters before and after amino acid-induced processing. Cultures of wild-type (WT; CAY60), asi1Δ (CAY150), asi1Δdal81Δ (MBY64), and dal81Δ (MBY79) strains carrying plasmid pMB10 (Stp1-HA) were grown in SD medium (− leu), and where indicated leucine was added 30 min prior to harvest (+ leu). Cell lysates were prepared and analyzed by ChIP using anti-HA antibody (3F10). Strain CAY60 (WT) carrying pCA029 (nontagged Stp1) was included to control nonspecific immunoprecipitation, and the ability to amplify the ACT1 promoter was assessed to control association with nonspecific DNA sequences. The size of amplified fragments are: AGP1, 246 bp; GNP1, 313 bp; and ACT1, 274 bp. The relative levels of immunoprecipitated promoter DNA are indicated (arbitrary units).