Proteomic analysis of extracellular ATP-regulated proteins identifies ATP synthase {beta}-subunit as a novel plant cell death regulator.

Chivasa, S; Tome, DF; Hamilton, JM; Slabas, AR.

Abstract

Abstract Extracellular ATP is an important signal molecule required to cue plant growth and developmental programmes, interactions with other organisms, and responses to environmental stimuli. The molecular targets mediating the physiological effects of extracellular ATP in plants have not yet been identified. We developed a well characterised experimental system that depletes Arabidopsis cell suspension culture extracellular ATP via treatment with the cell death-inducing mycotoxin fumonisin B1. This provided a platform for protein profile comparison between extracellular ATP-depleted cells and fumonisin B1-treated cells replenished with exogenous ATP, thus enabling the identification of proteins regulated by extracellular ATP signalling. Using two-dimensional difference in-gel electrophoresis and MALDI-TOF MS analysis of microsomal membrane and total soluble protein fractions, we identified 26 distinct proteins whose gene expression is controlled by the level of extracellular ATP. An additional 48 proteins that responded to fumonisin B1 were unaffected by extracellular ATP levels, confirming that this mycotoxin has physiological effects on Arabidopsis that are independent of its ability to trigger extracellular ATP depletion. Molecular chaperones, cellular redox control enzymes, glycolytic enzymes, and components of the cellular protein degradation machinery were among the extracellular ATP-responsive proteins. A major category of proteins highly regulated by extracellular ATP were components of ATP metabolism enzymes. We selected one of these, the mitochondrial ATP synthase β-subunit, for further analysis using reverse genetics. Plants in which the gene for this protein was knocked out by insertion of a transfer-DNA sequence became resistant to fumonisin B1-induced cell death. Therefore, in addition to its function in mitochondrial oxidative phosphorylation, our study defines a new role for ATP synthase β-subunit as a pro-cell death protein. More significantly, this protein is a novel target for extracellular ATP in its function as a key negative regulator of plant cell death.

Citation

Chivasa, S., Tome, D., Hamilton, J., & Slabas, A. (2010). Proteomic analysis of extracellular ATP-regulated proteins identifies ATP synthase {beta}-subunit as a novel plant cell death regulator

• Journal Article Type: Article
• Publication Date: 2010-12
• Deposit Date: Jan 12, 2011
• Journal: Molecular Cell Proteomics
• Publisher URL: http://www.ncbi.nlm.nih.gov/pubmed/21156838