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MOT1‐catalyzed TBP–DNA disruption: uncoupling DNA conformational change and role of upstream DNA
Author(s) -
Darst Russell P.,
Wang Dongyan,
Auble David T.
Publication year - 2001
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1093/emboj/20.8.2028
Subject(s) - atp hydrolysis , dna , biology , dna clamp , biophysics , atpase , helicase , walker motifs , biochemistry , microbiology and biotechnology , rna , enzyme , gene , reverse transcriptase
SNF2/SWI2‐related ATPases employ ATP hydrolysis to disrupt protein–DNA interactions, but how ATP hydrolysis is coupled to disruption is not understood. Here we examine the mechanism of action of MOT1, a yeast SNF2/SWI2‐related ATPase that uses ATP hydrolysis to remove TATA binding protein (TBP) from DNA. MOT1 function requires a 17 bp DNA ‘handle’ upstream of the TATA box, which must be double stranded. Remarkably, MOT1‐catalyzed disruption of TBP–DNA does not appear to require DNA strand separation, DNA bending or twisting of the DNA helix. Thus, TBP–DNA disruption is accomplished in a reaction apparently not driven by a change in DNA structure. MOT1 action is supported by DNA templates in which the handle is connected to the TATA box via single‐stranded DNA, indicating that the upstream duplex DNA can be conformationally uncoupled from the TATA box. Combining these results with proposed similarities between SNF2/SWI2 ATPases and helicases, we suggest that MOT1 uses ATP hydrolysis to translocate along the handle and thereby disrupt interactions between TBP and DNA.