Figure 1

Gal4-MOF activates expression of a UAS-RedStinger reporter gene. (A, B). Schematic representation of the constructs used in this study. The DNA binding domain [DB] of yeast Gal4 (aa 1-147) was fused in frame with a HA epitope tag and either full length MOF or MOF mutant. Expression was controlled with the constitutive hsp83 promoter. The numbers in parentheses correspond to the amino acid numbers in full length MOF, the mutations analysed in MOF are indicated, mutated amino acids are numbered. CD: chromo-related domain; Zn: CCHC zinc finger; HAT: acetyl transferase domain. (C) Gal4-MOF protein expression in adult flies was confirmed by western blotting with an anti-Gal4 antibody. Blots were stripped and re-probed with an alpha-tubulin antibody as a loading control. Similar blots were performed for all lines used in this study. (D) Schematic representation of the UAS-RedStinger reporter [28]. Expression of the DsRed.T4 red fluorescent protein is under the control of a minimal promoter from the hsp70 gene and upstream binding sites for Gal4 (UAS). The reporter is flanked by gypsy insulator elements (I). The nuclear localization sequence (nls) from TRA is fused to the C terminus of DsRed.T4. (E) Gal4-MOF activates UAS-RedStinger expression more strongly in females than males in larvae (top left panel) and adults (top right panel). DsRed.T4 fluorescence is highest in males that express high levels of Gal4-MOF protein such as line S41 (bottom right panel). Reporter line UAS-RedStinger4 (FBst0008546) responds more strongly to Gal4-MOF than line UAS-RedStinger6 (FBst0008547) (bottom right panel).