An E3 ubiquitin ligase from Brassica napus induces a typical heat-shock response in its own way in Escherichia coli

Acta Biochim Biophys Sin (Shanghai). 2017 Mar 1;49(3):262-269. doi: 10.1093/abbs/gmx004.

Abstract

Previously, we have identified a novel E3 ubiquitin ligase, BNTR1, which plays a key role in heat stress response in Brassica napus. In this study, we accidentally found that BNTR1 can also improve thermal tolerance and reduce growth inhibition at 42°C in Escherichia coli, in a manner different from that in plant. We show that BNTR1 activates E. coli heat-shock response at low concentration in soluble form instead of in inclusion body, but BNTR1 is not functioning as a heat-shock protein (HSP) because deficient temperature-sensitive mutants of HSP genes display unconspicuous thermal tolerance in the presence of BNTR1. Our further studies show that BNTR1 triggers heat-shock response by competing with σ32 (σ32, heat-shock transcription factor) to its binding proteins DnaJ (HSP40) and DnaK (HSP70), which results in the release and accumulation of σ32, thereby promoting the heat-shock response, even under the non-heat-shock conditions. At 37°C, accumulation of the HSPs induced by BNTR1 could make cells much more tolerant than those without BNTR1 at 42°C. Thus, our results suggest that BNTR1 may potentially be a promising target in fermentation industry for reducing impact from temperature fluctuation, where E. coli works as bioreactors.

Keywords: E3 ubiquitin ligase; heat-shock response; thermal resistance; σ32.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Blotting, Western
  • Brassica napus / enzymology*
  • Brassica napus / genetics
  • Cytoplasm / metabolism
  • Escherichia coli / genetics
  • Escherichia coli / metabolism*
  • Escherichia coli Proteins / metabolism*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • HSP40 Heat-Shock Proteins / metabolism
  • HSP70 Heat-Shock Proteins / metabolism
  • Heat-Shock Proteins / metabolism*
  • Heat-Shock Response / genetics
  • Microscopy, Fluorescence
  • Plant Proteins / genetics
  • Plant Proteins / metabolism*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Transformation, Genetic
  • Ubiquitin-Protein Ligases / genetics
  • Ubiquitin-Protein Ligases / metabolism*

Substances

  • DnaJ protein, E coli
  • Escherichia coli Proteins
  • HSP40 Heat-Shock Proteins
  • HSP70 Heat-Shock Proteins
  • Heat-Shock Proteins
  • Plant Proteins
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins
  • Ubiquitin-Protein Ligases
  • dnaK protein, E coli