The RING-Type E3 Ligase XBAT35.2 Is Involved in Cell Death Induction and Pathogen Response

Plant Physiol. 2017 Nov;175(3):1469-1483. doi: 10.1104/pp.17.01071. Epub 2017 Sep 26.

Abstract

XBAT35 belongs to a subfamily of Arabidopsis (Arabidopsis thaliana) RING-type E3s that are similar in domain architecture to the rice (Oryza sativa) XA21 Binding Protein3, a defense protein. The XBAT35 transcript undergoes alternative splicing to produce two protein isoforms, XBAT35.1 and XBAT35.2. Here, we demonstrate that XBAT35.2 localizes predominantly to the Golgi and is involved in cell death induction and pathogen response. XBAT35.2, but not XBAT35.1, was found to trigger cell death when overexpressed in tobacco (Nicotiana benthamiana) leaves and does so in a manner that requires its RING domain. Loss of XBAT35 gene function disrupts the plant's ability to defend against pathogen attack, whereas overexpression of XBAT35.2 enhances resistance to pathogens. XBAT35.2 was found to be unstable and promotes its own degradation, suggesting self-regulation. Inoculation with virulent and avirulent strains of the bacterial pathogen Pseudomonas syringae pv tomato DC3000 results in a drastic reduction in the levels of ubiquitinated XBAT35.2 and an increase in the abundance of the E3. This implies that pathogen infection prohibits XBAT35.2 self-regulation and stabilizes the E3. In agreement with a role in defending against pathogens, XBAT35.2 interacts with defense-related Accelerated Cell Death11 (ACD11) in planta and promotes the proteasome-dependent turnover of ACD11 in cell-free degradation assays. In accordance with regulation by a stabilized XBAT35.2, the levels of ubiquitinated ACD11 increased considerably, and the abundance of ACD11 was reduced following pathogen infection. In addition, treatment of transgenic seedlings with a proteasome inhibitor results in the accumulation of ACD11, confirming proteasome-dependent degradation. Collectively, these results highlight a novel role for XBAT35.2 in cell death induction and defense against pathogens.

MeSH terms

  • Arabidopsis / cytology*
  • Arabidopsis / genetics
  • Arabidopsis / microbiology*
  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / metabolism*
  • Cell Death
  • Disease Resistance
  • Golgi Apparatus / metabolism
  • Plant Cells / metabolism
  • Plant Diseases / microbiology
  • Plant Leaves / metabolism
  • Plants, Genetically Modified
  • Proteasome Endopeptidase Complex / metabolism
  • Protein Binding
  • Proteolysis
  • Pseudomonas syringae / pathogenicity
  • Pseudomonas syringae / physiology*
  • RING Finger Domains
  • Subcellular Fractions / metabolism
  • Tobacco / cytology
  • Tobacco / genetics
  • Ubiquitin-Protein Ligases / genetics
  • Ubiquitin-Protein Ligases / metabolism*
  • Ubiquitination
  • Virulence

Substances

  • Arabidopsis Proteins
  • Ubiquitin-Protein Ligases
  • XBAT35 protein, Arabidopsis
  • Proteasome Endopeptidase Complex