FIGURE 11:

The Hot1 protein binds the HoRE of the STL1 promoter. (A) The indicated quantities of recombinant histidine (His)-tagged Hot1 protein were mixed with ³²P-labeled fragment of the STL1 promoter for 15 min and then loaded onto 5% acrylamide gel. Gels were run at 110 V for 6 h. Recombinant His-tagged JNK2, BSA, or GST protein purified from E. coli were tested as controls. (B) Recombinant His-tagged Hot1 (1 μg) was mixed with ³²P-labeled fragment together with the indicated unlabeled fragments. (C) The ³²P-labeled fragment was incubated with cell lysates prepared from wild-type or hot1∆ cells exposed or not exposed to 0.7 M NaCl. (D) Cell lysates of the indicated strains were incubated with ³²P-labeled E3 fragment of the STL1 promoter (see Figure 7; see sequence at bottom) or with a mutated E3 fragment (M; see sequence at bottom).