Tetrameric architecture of an active phenol-bound form of the AAA+ transcriptional regulator DmpR

Nat Commun. 2020 Jun 1;11(1):2728. doi: 10.1038/s41467-020-16562-5.

Abstract

The Pseudomonas putida phenol-responsive regulator DmpR is a bacterial enhancer binding protein (bEBP) from the AAA+ ATPase family. Even though it was discovered more than two decades ago and has been widely used for aromatic hydrocarbon sensing, the activation mechanism of DmpR has remained elusive. Here, we show that phenol-bound DmpR forms a tetramer composed of two head-to-head dimers in a head-to-tail arrangement. The DmpR-phenol complex exhibits altered conformations within the C-termini of the sensory domains and shows an asymmetric orientation and angle in its coiled-coil linkers. The structural changes within the phenol binding sites and the downstream ATPase domains suggest that the effector binding signal is propagated through the coiled-coil helixes. The tetrameric DmpR-phenol complex interacts with the σ54 subunit of RNA polymerase in presence of an ATP analogue, indicating that DmpR-like bEBPs tetramers utilize a mechanistic mode distinct from that of hexameric AAA+ ATPases to activate σ54-dependent transcription.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases / chemistry*
  • Adenosine Triphosphatases / genetics
  • Adenosine Triphosphatases / metabolism
  • Adenosine Triphosphate / metabolism
  • Amino Acid Sequence
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Binding Sites / genetics
  • DNA-Binding Proteins / chemistry*
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • DNA-Directed RNA Polymerases / metabolism
  • Gene Expression Regulation, Bacterial
  • Phenol / metabolism
  • Protein Binding
  • Protein Conformation*
  • Protein Multimerization*
  • Pseudomonas putida / enzymology
  • Pseudomonas putida / genetics
  • Sequence Homology, Amino Acid
  • Trans-Activators / chemistry*
  • Trans-Activators / genetics
  • Trans-Activators / metabolism

Substances

  • Bacterial Proteins
  • DNA-Binding Proteins
  • DmpR protein, Pseudomonas
  • Trans-Activators
  • Phenol
  • Adenosine Triphosphate
  • DNA-Directed RNA Polymerases
  • Adenosine Triphosphatases