Interaction of TIP26 from a hyperthermophilic archaeon with TFB/TBP/DNA ternary complex.
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.
Matsuda
T
T
Department of Material and Life Science, Graduate School of Engineering, Osaka University, Japan.
Fujikawa
M
M
Haruki
M
M
Tang
X F
XF
Ezaki
S
S
Imanaka
T
T
Morikawa
M
M
Kanaya
S
S
D000595:Amino Acid Sequence; D019843:Archaeal Proteins; D003001:Cloning, Molecular; D019641:DNA, Archaeal / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D019766:Genes, Archaeal; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D035181:TATA-Box Binding Protein; D013696:Temperature; D019712:Thermococcus / Q000201:enzymology / Q000235:genetics; D035581:Transcription Factor TFIIB; D014157:Transcription Factors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D019843:Archaeal Proteins; D019641:DNA, Archaeal; D004268:DNA-Binding Proteins; D046911:Macromolecular Substances; D009687:Nuclear Proteins; D011994:Recombinant Proteins; D035181:TATA-Box Binding Protein; C121585:TIP protein, Pyrococcus kodakaraensis; D035581:Transcription Factor TFIIB; D014157:Transcription Factors; C106466:transcription factor B, Archaea