Interaction of TIP26 from a hyperthermophilic archaeon with TFB/TBP/DNA ternary complex |
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Authors: | Tomoki Matsuda Makoto Fujikawa Mitsuru Haruki Xiao-Feng Tang Satoshi Ezaki Tadayuki Imanaka Masaaki Morikawa Shigenori Kanaya |
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Institution: | Department of Material and Life Science, Graduate School of Engineering, Osaka University, Japan. |
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Abstract: | Interactions of TBP-interacting protein (TIP26), TBP, and TFB from a hyperthermophilic archaeon Thermococcus kodakaraensis KOD1 with TATA-DNA were examined by electrophoretic mobility shift assay. Tk-TFB formed a ternary complex with Tk-TBP and TATA-DNA. Tk-TIP26 did not inhibit the formation of this ternary complex, but interacted with it to form a TIP26/TFB/TBP/DNA quaternary complex. This interaction is rather weak, and a large excess of Tk-TIP26 over Tk-TBP is required to fully convert the TFB/TBP/DNA ternary complex to the quaternary complex. However, determination of the concentration of Tk-TIP26 and Tk-TBP in KOD1 cells by Western blotting analysis indicated that the concentration of Tk-TIP26 is approximately ten times that of Tk-TBP, suggesting that the quaternary complex might also form in vivo. |
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