Structures of regulatory machinery reveal novel molecular mechanisms controlling B. subtilis nitrogen homeostasis

Genes Dev. 2015 Feb 15;29(4):451-64. doi: 10.1101/gad.254714.114.

Abstract

All cells must sense and adapt to changing nutrient availability. However, detailed molecular mechanisms coordinating such regulatory pathways remain poorly understood. In Bacillus subtilis, nitrogen homeostasis is controlled by a unique circuitry composed of the regulator TnrA, which is deactivated by feedback-inhibited glutamine synthetase (GS) during nitrogen excess and stabilized by GlnK upon nitrogen depletion, and the repressor GlnR. Here we describe a complete molecular dissection of this network. TnrA and GlnR, the global nitrogen homeostatic transcription regulators, are revealed as founders of a new structural family of dimeric DNA-binding proteins with C-terminal, flexible, effector-binding sensors that modulate their dimerization. Remarkably, the TnrA sensor domains insert into GS intersubunit catalytic pores, destabilizing the TnrA dimer and causing an unprecedented GS dodecamer-to-tetradecamer conversion, which concomitantly deactivates GS. In contrast, each subunit of the GlnK trimer "templates" active TnrA dimers. Unlike TnrA, GlnR sensors mediate an autoinhibitory dimer-destabilizing interaction alleviated by GS, which acts as a GlnR chaperone. Thus, these studies unveil heretofore unseen mechanisms by which inducible sensor domains drive metabolic reprograming in the model Gram-positive bacterium B. subtilis.

Keywords: MerR family; TnrA/GlnR family; dodecamer; glutamine synthetase; tetradecamer.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacillus subtilis / genetics*
  • Bacillus subtilis / metabolism*
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Crystallization
  • DNA / chemistry
  • DNA / metabolism
  • Dimerization
  • Enzyme Activation / genetics
  • Glutamate-Ammonia Ligase / chemistry
  • Glutamate-Ammonia Ligase / metabolism
  • Homeostasis / genetics*
  • Models, Molecular*
  • Nitrogen / metabolism*
  • Protein Structure, Tertiary
  • Repressor Proteins / metabolism
  • Sequence Alignment

Substances

  • Bacterial Proteins
  • Repressor Proteins
  • ScgR protein, Bacillus subtilis
  • DNA
  • Glutamate-Ammonia Ligase
  • Nitrogen