Figure 8.

Functional complementation with HK1b mutants. (A) Schematic representation of HK1b with the different domains as indicated. All mutated positions are indicated with an asterisk and the corresponding mutated sequences shown on the right. The putative canonical histidine and aspartate residues are in bold; (B) Wild-type and single mutation constructs of HK1b were introduced into the osmodeficient strain (MH179) as indicated and spotted onto different media at a density corresponding to an OD₆₀₀ = 1 and three 10-fold dilutions as indicated. The resulting transformants were spotted onto SG-Ura medium for growth control and onto SD-Ura medium for growth rescue. Finally, transformants were spotted onto SD-Ura+NaCl media with increasing concentrations of NaCl for the osmotic tolerance test; (C) Double and triple mutation constructs of HK1b were introduced into the osmodeficient strain (MH179) as indicated and spotted onto different media at a density corresponding to an OD₆₀₀ = 1 and three 10-fold dilutions as indicated. The resulting transformants were spotted onto SG-Ura medium for the growth control and onto SD-Ura medium for growth rescue. Finally, transformants were spotted onto SD-Ura+NaCl medium with 0.9 M of NaCl for the osmotic tolerance test.