Subunit-selective proteasome activity profiling uncovers uncoupled proteasome subunit activities during bacterial infections

Plant J. 2017 Apr;90(2):418-430. doi: 10.1111/tpj.13494. Epub 2017 Mar 14.

Abstract

The proteasome is a nuclear-cytoplasmic proteolytic complex involved in nearly all regulatory pathways in plant cells. The three different catalytic activities of the proteasome can have different functions, but tools to monitor and control these subunits selectively are not yet available in plant science. Here, we introduce subunit-selective inhibitors and dual-color fluorescent activity-based probes for studying two of the three active catalytic subunits of the plant proteasome. We validate these tools in two model plants and use this to study the proteasome during plant-microbe interactions. Our data reveal that Nicotiana benthamiana incorporates two different paralogs of each catalytic subunit into active proteasomes. Interestingly, both β1 and β5 activities are significantly increased upon infection with pathogenic Pseudomonas syringae pv. tomato DC3000 lacking hopQ1-1 [PtoDC3000(ΔhQ)] whilst the activity profile of the β1 subunit changes. Infection with wild-type PtoDC3000 causes proteasome activities that range from strongly induced β1 and β5 activities to strongly suppressed β5 activities, revealing that β1 and β5 activities can be uncoupled during bacterial infection. These selective probes and inhibitors are now available to the plant science community, and can be widely and easily applied to study the activity and role of the different catalytic subunits of the proteasome in different plant species.

Keywords: Arabidopsis thaliana; Nicotiana benthamiana; activity-based protein profiling; catalytic subunit; core protease; proteasome manipulation; technical advance.

MeSH terms

  • Arabidopsis / metabolism*
  • Arabidopsis / microbiology*
  • Bacterial Infections / metabolism*
  • Plant Diseases / microbiology*
  • Proteasome Endopeptidase Complex / metabolism*
  • Pseudomonas syringae / pathogenicity*
  • Tobacco / metabolism*
  • Tobacco / microbiology*

Substances

  • Proteasome Endopeptidase Complex