The chimeric cyclic nucleotide-gated ion channel ATCNGC11/12 constitutively induces programmed cell death in a Ca2+ dependent manner

Plant Mol Biol. 2007 Dec;65(6):747-61. doi: 10.1007/s11103-007-9239-7. Epub 2007 Sep 21.

Abstract

The hypersensitive response (HR) involves programmed cell death (PCD) in response to pathogen infection. To investigate the pathogen resistance signaling pathway, we previously identified the Arabidopsis mutant cpr22, which displays constitutive activation of multiple defense responses including HR like cell death. The cpr22 mutation has been identified as a 3 kb deletion that fuses two cyclic nucleotide-gated ion channel (CNGC)-encoding genes, ATCNGC11 and ATCNGC12, to generate a novel chimeric gene, ATCNGC11/12. In this study, we conducted a characterization of cell death induced by transient expression of ATCNGC11/12 in Nicotiana benthamiana. Electron microscopic analysis of this cell death showed similar characteristics to PCD, such as plasma membrane shrinkage and vesicle formation. The hallmark of animal PCD, fragmentation of nuclear DNA, was also observed in ATCNGC11/12-induced cell death. The development of cell death was significantly suppressed by caspase-1 inhibitors, suggesting the involvement of caspases in this process. Recently, vacuolar processing enzyme (VPE) was isolated as the first plant caspase-like protein, which is involved in HR development. In VPE-silenced plants development of cell death induced by ATCNGC11/12 was much slower and weaker compared to control plants, suggesting the involvement of VPE as a caspase in ATCNGC11/12-induced cell death. Complementation analysis using a Ca2+ uptake deficient yeast mutant demonstrated that the ATCNGC11/12 channel is permeable to Ca2+. Additionally, calcium channel blockers such as GdCl3 inhibited ATCNGC11/12-induced HR formation, whereas potassium channel blockers did not. Taken together, these results indicate that the cell death that develops in the cpr22 mutant is indeed PCD and that the chimeric channel, ATCNGC11/12, is at the point of, or up-stream of the calcium signal necessary for the development of HR.

Publication types

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

MeSH terms

  • Apoptosis / drug effects
  • Apoptosis / physiology*
  • Arabidopsis / cytology
  • Arabidopsis / metabolism
  • Arabidopsis Proteins / analysis
  • Arabidopsis Proteins / genetics*
  • Arabidopsis Proteins / physiology
  • Calcium Channels / analysis
  • Calcium Channels / genetics*
  • Calcium Channels / physiology
  • Calcium Signaling*
  • Caspase Inhibitors
  • Caspases / physiology
  • Cyclic Nucleotide-Gated Cation Channels / analysis
  • Cyclic Nucleotide-Gated Cation Channels / genetics*
  • Cyclic Nucleotide-Gated Cation Channels / physiology
  • Cysteine Endopeptidases / physiology
  • Cysteine Proteinase Inhibitors / pharmacology
  • DNA Fragmentation
  • Mutant Chimeric Proteins / analysis
  • Mutant Chimeric Proteins / physiology*
  • Plants, Genetically Modified / cytology
  • Plants, Genetically Modified / drug effects
  • Plants, Genetically Modified / metabolism
  • Tobacco / genetics

Substances

  • Arabidopsis Proteins
  • CNGC11 protein, Arabidopsis
  • CNGC12 protein, Arabidopsis
  • Calcium Channels
  • Caspase Inhibitors
  • Cyclic Nucleotide-Gated Cation Channels
  • Cysteine Proteinase Inhibitors
  • Mutant Chimeric Proteins
  • Caspases
  • Cysteine Endopeptidases
  • vacuolar processing enzyme