Figure 3

Interactions between MBD2/3 and NuRD homologues in a yeast two-hybrid assay (A) An MBD2/3 long isoform bait efficiently transactivated reporter gene transcription in yeast expressing p55 or MBD2/3 small isoform prey constructs, respectively. A weaker transactivation could be seen upon expression of a MI-2 prey construct (panel M). An MBD2/3 small isoform bait transactivated reporter gene transcription co-expressing a MI-2 prey also under high-stringent conditions (panel H). No transactivation could be observed with MTA-like or RPD3 preys. N shows growth on non-selective plates, M on medium-stringency plates and H on high-stringency plates. X indicates X-gal staining of colonies grown on non-selective plates. (B and C) Delineation of interaction domains in a yeast two-hybrid assay. The methyl-DNA binding domain (MBD) is highlighted in red, the Drosophila specific domain (DSD) in green and the C-terminal coiled-coil domain (cc) in yellow. (B) The C-terminal region of MBD2/3 interacts with p55. The full-length p55 construct was used as a bait and various deletion mutants of MBD2/3 were used as preys. This identified the region between amino acids 178 and 305 of the MBD2/3 long isoform as the p55 interaction domain. (C) Deletion of the coiled-coil domain abolishes interactions between MBD2/3 small isoform and MI-2 under high stringent conditions.