Fig. 6.

Interactions among T. brucei RAD51 paralogues and with RAD51. A. Yeast two-hybrid analysis of interactions. β-Galactosidase activity is shown for yeast cells carrying plasmids that express RAD51, RAD51-3, RAD51-4, RAD51-5 or RAD51-6 fused with a LexA DNA binding domain (LexA), in combination with plasmids expressing any of the proteins fused with a B42 transcriptional activation domain (B42); activity is also shown for control yeast cells carrying the LexA fusion-expressing plasmids and the B42 vector without a T. brucei gene (Empty). Values are the means of three independent experiments for two independent co-transformant yeast clones; deviation in activity around the mean is indicated (ND, not determined). B. Interactions between a GST–RAD51-3 fusion coexpressed in E. coli with native RAD51-6, or with a GST–RAD51-4 fusion coexpressed with native RAD51-3 or RAD51-6. Western blots (anti-RAD51 paralogue antiserum used is indicated by WB) are shown of the native proteins in cell extracts before purification (input), and after GST pull down of the GST fusion proteins from the cell extract (eluate) using glutathione beads. Controls are shown with GST fusion proteins expressed on their own, and with native RAD51 paralogues expressed with GST alone. Expression of the GST fusions, or of GST, by western analysis is not shown.