ATP hydrolysis-dependent formation of a dynamic ternary nucleoprotein complex with MutS and MutL.
Functional interactions of Escherichia coli MutS and MutL in mismatch repair are dependent on ATP. In this study, we show that MutS and MutL associate with immobilised DNA in a manner dependent on ATP hydrolysis and with an ATP concentration near the solution K m of the ATPase of MutS. After removal of MutS, MutL and ATP, much of the protein in this ternary complex is not stably associated, with MutL leaving the complex more rapidly than MutS. The rapid dissociation reveals a dynamic interaction with concurrent rapid association and dissociation of proteins from the DNA. Analysis by surface plasmon resonance showed that the DNA interacting with dynamically bound protein was more resistant to nuclease digestion than the DNA in MutS-DNA complexes. Non-hydrolysable analogs of ATP inhibit the formation of this dynamic complex, but permit formation of a second type of ternary complex with MutS and MutL stably bound to the immobilised DNA.
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D020137:Base Pair Mismatch; D004268:DNA-Binding Proteins; D003850:Deoxyribonuclease I / Q000378:metabolism; D029968:Escherichia coli Proteins; D006801:Humans; D006868:Hydrolysis; D000070956:MutL Proteins; D051719:MutS DNA Mismatch-Binding Protein; D009698:Nucleoproteins / Q000378:metabolism*; D020349:Surface Plasmon Resonance