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Effect of Gal80 on the phosphorylation of two substrates. Srb10/11 complex was incubated with 0.1 nmol of either Gst-Gal4 (683–881) or Gst-CTD3-Gal4 (840–881) in the presence or absence of 2-fold molar excess of Gal80. We expect that Gal80 binds stoichiometrically to the Gal4-activating region (840–874) under these conditions (17). We use three repeats of the CTD heptapeptide YSPTSPS as a substrate based on experiments of Hengartner et al. (8).
