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. 2004 Feb 1;18(3):333–343. doi: 10.1101/gad.1148404

Figure 5.

Figure 5.

The Gal4-Tra1 interaction is required for SAGA recruitment and transcriptional activation. (A) Fluorescence emission spectra of yeast strains expressing Tra1-EYFP and ECFP fused to either Gal4 or derivatives lacking its AD, Gal4(ΔAD)-ECFP, or its DNA-binding domain, Gal4(ΔDBD)-ECFP. (B) Transcriptional analysis of GAL1 and SED1 by primer-extension (left) and ChIP analysis of Gal4 and SAGA recruitment to the GAL1 UAS (right) following inactivation of Tra1. Formaldehyde-based in vivo cross-linking and ChIP was performed as previously described (Bhaumik and Green 2001). Immunoprecipitated (IP) DNA was amplified by PCR using primer pairs corresponding to the GAL1 UAS, and IP DNA was quantitated and presented as the ratio of IP to input relative to wild type. (C) Fluorescence emission spectra in Gal4-ECFP/Tra1-EYFP cells in the presence and absence of the SAGA subunit Spt20. (Inset) Immunoblot showing Tra1-EYFP levels in wild-type and spt20Δ strains. (D) ChIP analysis of Tra1 recruitment to the GAL1 UAS in wild-type cells and in an spt20Δ deletion mutant.