title: Plant programmed cell death caused by an autoactive form of Prf is suppressed by co-expression of the Prf LRR domain.
abstract: In tomato, the NBARC-LRR resistance (R) protein Prf acts in concert with the Pto or Fen kinase to determine immunity against Pseudomonas syringae pv. tomato (Pst). Prf-mediated defense signaling is initiated by the recognition of two sequence-unrelated Pst-secreted effector proteins, AvrPto and AvrPtoB, by tomato Pto or Fen. Prf detects these interactions and activates signaling leading to host defense responses including localized programmed cell death (PCD) that is associated with the arrest of Pst growth. We found that Prf variants with single amino acid substitutions at D1416 in the IHD motif (isoleucine-histidine-aspartic acid) in the NBARC domain cause effector-independent PCD when transiently expressed in leaves of Nicotiana benthamiana, suggesting D1416 plays an important role in activation of Prf. The N-terminal region of Prf (NPrf) and the LRR domain are required for this autoactive Prf cell death signaling but dispensable for accumulation of the Prf(D1416V) protein. Significantly, co-expression of the Prf LRR but not NPrf, with Prf(D1416V), AvrPto/Pto, AvrPtoB/Pto, an autoactive form of Pto (Pto(Y207D)), or Fen completely suppresses PCD. However, the Prf LRR does not interfere with PCD caused by Rpi-blb1(D475V), a distinct R protein-mediated PCD signaling event, or that caused by overexpression of MAPKKKα, a protein acting downstream of Prf. Furthermore, we found the Prf(D1416V) protein is unable to accumulate in plant cells when co-expressed with the Prf LRR domain, likely explaining the cell death suppression. The mechanism for the LRR-induced degradation of Prf(D1416V) is unknown but may involve interference in the intramolecular interactions of Prf or to binding of the unattached LRR to other host proteins that are needed for Prf stability.
authors.affiliation: Department of Plant, Soil and Entomological Sciences, University of Idaho, Moscow, ID 83844, USA.
authors.forename: Xinran
authors.initials: X
authors.lastname: Du
authors.forename: Min
authors.initials: M
authors.lastname: Miao
authors.forename: Xinrong
authors.initials: X
authors.lastname: Ma
authors.forename: Yongsheng
authors.initials: Y
authors.lastname: Liu
authors.forename: Joseph C
authors.initials: JC
authors.lastname: Kuhl
authors.forename: Gregory B
authors.initials: GB
authors.lastname: Martin
authors.forename: Fangming
authors.initials: F
authors.lastname: Xiao
mesh_terms: D020816:Amino Acid Motifs; D017209:Apoptosis; D016923:Cell Death; D015536:Down-Regulation; D015870:Gene Expression; D018506:Gene Expression Regulation, Plant; D018551:Solanum lycopersicum / Q000235:genetics / Q000378:metabolism / Q000382:microbiology; D010935:Plant Diseases / Q000382:microbiology*; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D044224:Pseudomonas syringae / Q000502:physiology; D015398:Signal Transduction; D014026:Nicotiana / Q000166:cytology* / Q000378:metabolism / Q000382:microbiology
chemical_list: D010940:Plant Proteins