Figure 3

Schematic overview of the MBF1 from yeast, T. tenax and M. mazei , and constructed chimeric Archaea/yeast MBF1 proteins. (A) Chimeric proteins were generated by the use of recombination/gap repair cloning technique and the vector pRS316 under control of the natural yeast MBF1 regulatory region. (B) Functional complementation of yeast deletion mutant by yeast Mbf1 (y), aMBF1 (T. tenax (T) and M. mazei (M)), and chimeric MBF1 variants. Only mbf1Δ expressing MBF1 comprising the C-terminal extension of T. tenax (yyT) or M. mazei (yyM) MBF1 restored WT activity, i.e. AT resistance. The yyΔCt yMBF1 mutant lacking the C-terminal part (residues 138-151) was used as control. The assay was performed as described above (see Figure 2).