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. 2006 Aug 29;398(Pt 3):461–467. doi: 10.1042/BJ20060406

Figure 4. The Not-Box of CNOT2 interacts with different subunits of the SMRT/NCoR–HDAC3 complex in a yeast two-hybrid assay.

Figure 4

(A) Schematic drawing showing the known functional domains of SMRT and NCoR proteins. (B) The Not-Box of CNOT2 interacts with RD4 and with the RID of SMRT and NCoR. EGY48 cells were transformed with LexA and B42 expression plasmids together with a reporter plasmid. Galactosidase activity was detected by X-Gal (5-bromo-4-chloroindol-3-yl β-D-galactopyranoside) staining of yeast cells. The LexA fusion proteins are indicated to the left and the B42 fusion proteins are indicated above the panel. Full-length CNOT2 could not be tested as the LexA fusion protein behaves as a strong transcriptional activator in this assay. (C) Quantification of β-galactosidase activity. Lysates of yeast cells were prepared as described in the Experimental section. The indicated fold induction is relative to the activity of yeast cells expressing LexA and B42 control. (D) The Not-Box of CNOT2 interacts with GPS2 but not HDAC3 in the yeast two-hybrid assay. The assay was performed and the interactions were quantified as in (B, C).