Figure 1

Identification of transcriptional activating domain at the amino-terminus of zebrafish ZNF143. (A) Primary structure of zebrafish ZNF143. Numbers on top depict the amino acids at the beginning and end of prominent primary structure features of the protein. Numbers in parentheses are percentages of identical amino acid residues between the zebrafish and human proteins for various regions of primary structure. (B) HEK293 cells were transfected with 200 ng of pGL3/G5AdMLT firefly luciferase reporter plasmid, 10 ng of the particular pCI/GAL4DBD/zZNF143 effector plasmid DNA noted in the figure, and 200 ng of pRL-SV40 renilla luciferase reporter. The fold-activation was determined by comparing the firefly luciferase/renilla luciferase ratios for each sample to that ratio for samples where no effector plasmid was added. Bar height shows the mean number from different transfected samples (number of experiments reported in parentheses), and the error bar represents the standard deviation from the mean. Double asterisks signify p-value <0.01, and single asterisks signify a p-value <0.05 relative to samples transfected with GAL4DBD only. The panel below shows the relative expression of each GAL4DBD/zZNF143 fusion protein in HEK293 cells by immunoblot analysis using anti-GAL4DBD antibodies. (C) Zebrafish ZF4 cells were transfected, and relative expression levels analyzed exactly as described for (B). We were unable to detect GAL4DBD/zZNF143 fusion proteins in transfected ZF4 cells, possibly because of the much lower transfection efficiency compared to HEK293 cells.