Elucidation of the Features of the Zinc Finger Associated Domain (ZAD) Family of Transcription Factors in Drosophila Melanogaster

The zinc finger associated domain (ZAD) containing family of transcription factors is not well described in the literature, in part because it is very difficult to study by mutagenesis. We used in vitro -binding techniques to identify characteristics of the ZAD family, by constructing glutathione S-transferase (GST)-ZAD domain chimeric proteins for use in protein binding assays, and GST-Zinc finger array domain chimera for binding site selections. Protein binding assays indicated a possible shared cofactor, as seen in the analogous KRAB system in mammals. DNA binding assays have provided a consensus binding sequence for five of the ZAD proteins, consistent with previously reported work on ZAD and unpublished work on mammalian transcription factors. Research is ongoing with an additional ∼50 ZAD proteins to more fully map the binding characters of ZAD proteins.