Regulation of reactive oxygen species during plant immunity through phosphorylation and ubiquitination of RBOHD

Nat Commun. 2020 Apr 15;11(1):1838. doi: 10.1038/s41467-020-15601-5.

Abstract

Production of reactive oxygen species (ROS) is critical for successful activation of immune responses against pathogen infection. The plant NADPH oxidase RBOHD is a primary player in ROS production during innate immunity. However, how RBOHD is negatively regulated remains elusive. Here we show that RBOHD is regulated by C-terminal phosphorylation and ubiquitination. Genetic and biochemical analyses reveal that the PBL13 receptor-like cytoplasmic kinase phosphorylates RBOHD's C-terminus and two phosphorylated residues (S862 and T912) affect RBOHD activity and stability, respectively. Using protein array technology, we identified an E3 ubiquitin ligase PIRE (PBL13 interacting RING domain E3 ligase) that interacts with both PBL13 and RBOHD. Mimicking phosphorylation of RBOHD (T912D) results in enhanced ubiquitination and decreased protein abundance. PIRE and PBL13 mutants display higher RBOHD protein accumulation, increased ROS production, and are more resistant to bacterial infection. Thus, our study reveals an intricate post-translational network that negatively regulates the abundance of a conserved NADPH oxidase.

Publication types

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

MeSH terms

  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / metabolism*
  • Gene Expression Regulation, Plant / genetics
  • Gene Expression Regulation, Plant / physiology
  • NADPH Oxidases / genetics
  • NADPH Oxidases / metabolism*
  • Phosphorylation / genetics
  • Phosphorylation / physiology
  • Plant Diseases / genetics
  • Plant Immunity / genetics
  • Plant Immunity / physiology*
  • Protein Domains / genetics
  • Protein Domains / physiology
  • Protein Serine-Threonine Kinases / genetics
  • Protein Serine-Threonine Kinases / metabolism*
  • Reactive Oxygen Species / metabolism*
  • Signal Transduction / genetics
  • Signal Transduction / physiology
  • Ubiquitination / genetics
  • Ubiquitination / physiology

Substances

  • Arabidopsis Proteins
  • Reactive Oxygen Species
  • respiratory burst oxidase homolog D, Arabidopsis
  • NADPH Oxidases
  • PBL13 protein, Arabidopsis
  • Protein Serine-Threonine Kinases