Figure 2.
ATX1 Interacts with the CTD of Pol II.
Yeast two-hybrid or in vitro interaction assays were performed.
(A) The different regions of ATX1 containing the indicated domains (PWWP, DAST, ePHD, and SET) that were tested in yeast two-hybrid analyses are shown.
(B) The DNA binding domain (BD) or BD-CTD fusions (CTD) were tested for binding to the activation domain (AD) fused to the various ATX1 domains shown in (A). The growth of two dilutions (4 × 10−3 and 8 × 10−4) of the yeast culture on SD medium lacking Trp, Leu, His, and adenine is shown.
(C) and (D) Representative immunoblots of the input amounts of a soluble protein or the amount of this protein bound to beads containing a surface-bound control tag or a tag fusion protein. The soluble protein being tested is denoted at the top of each panel.
(C) Beads containing a His tag (His) or a His tag fused to the CTD of Pol II (His-CTD) were assayed for their ability to bind a soluble GST fusion to the SET domain of ATX1 (GST-SET). Input or bound protein was detected with antibody to GST (Anti GST).
(D) Beads containing a GST tag or a GST tag fused to the SET domain of ATX1 (GST-SET) were assayed for their ability to bind a soluble CTD of Pol II fused to a His tag (His-CTD) and detected with antibody to the CTD of Pol II (Anti CTD). All experiments were repeated three times.