Formation of MgF3 (-)-dependent complexes between an AAA(+) ATPase and σ(54.).

The widely distributed bacterial σ(54)-dependent transcription regulates pathogenicity and numerous adaptive responses in diverse bacteria. Formation of the σ(54)-dependent open promoter complex is a multi-step process driven by AAA(+) ATPases. Non-hydrolysable nucleotide analogues are particularly suitable for studying such complexity by capturing various intermediate states along the energy coupling pathway. Here we report a novel ATP analogue, ADP-MgF3 (-), which traps an AAA(+) ATPase with its target σ(54). The MgF3 (-)-dependent complex is highly homogeneous and functional assays suggest it may represent an early transcription intermediate state valuable for structural studies.