Acyl-CoA-binding protein 2 binds lysophospholipase 2 and lysoPC to promote tolerance to cadmium-induced oxidative stress in transgenic Arabidopsis

Plant J. 2010 Jun 1;62(6):989-1003. doi: 10.1111/j.1365-313X.2010.04209.x. Epub 2010 Mar 19.

Abstract

Lysophospholipids are intermediates of phospholipid metabolism resulting from stress and lysophospholipases detoxify lysophosphatidylcholine (lysoPC). Many lysophospholipases have been characterized in mammals and bacteria, but few have been reported from plants. Arabidopsis thaliana lysophospholipase 2 (lysoPL2) (At1g52760) was identified as a protein interactor of acyl-CoA-binding protein 2 (ACBP2) in yeast two-hybrid analysis and co-immunoprecipitation assays. BLASTP analysis indicated that lysoPL2 showed approximately 35% amino acid identity to the lysoPL1 family. Co-localization of autofluorescence-tagged lysoPL2 and ACBP2 by confocal microscopy in agroinfiltrated tobacco suggests the plasma membrane as a site for their subcellular interaction. LysoPL2 mRNA was induced by zinc (Zn) and hydrogen peroxide (H(2)O(2)), and lysoPL2 knockout mutants showed enhanced sensitivity to Zn and H(2)O(2) in comparison to wild type. LysoPL2-overexpressing Arabidopsis was more tolerant to H(2)O(2) and cadmium (Cd) than wild type, suggesting involvement of lysoPL2 in phospholipid repair following lipid peroxidation arising from metal-induced stress. Lipid hydroperoxide (LOOH) contents in ACBP2-overexpressors and lysoPL2-overexpressors after Cd-treatment were lower than wild type, indicating that ACBP2 and lysoPL2 confer protection during oxidative stress. A role for lysoPL2 in lysoPC detoxification was demonstrated when recombinant lysoPL2 was observed to degrade lysoPC in vitro. Filter-binding assays and Lipidex competition assays showed that (His)(6)-ACBP2 binds lysoPC in vitro. Binding was disrupted in a (His)(6)-ACBP2 derivative lacking the acyl-CoA-binding domain, confirming that this domain confers lysoPC binding. These results suggest that ACBP2 can bind both lysoPC and lysoPL2 to promote the degradation of lysoPC in response to Cd-induced oxidative stress.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Arabidopsis / enzymology*
  • Arabidopsis / genetics
  • Arabidopsis Proteins / metabolism*
  • Cadmium / adverse effects*
  • Carrier Proteins / metabolism*
  • Cell Membrane / metabolism
  • Gene Expression Regulation, Plant
  • Gene Knockout Techniques
  • Lipid Peroxidation
  • Lysophospholipase / genetics
  • Lysophospholipase / metabolism*
  • Molecular Sequence Data
  • Mutagenesis, Insertional
  • Mutation
  • Oxidative Stress*
  • Plants, Genetically Modified / genetics
  • Plants, Genetically Modified / metabolism
  • Protein Binding
  • RNA, Plant / genetics
  • Stress, Physiological
  • Two-Hybrid System Techniques

Substances

  • ACBP2 protein, Arabidopsis
  • Arabidopsis Proteins
  • Carrier Proteins
  • RNA, Plant
  • Cadmium
  • Lysophospholipase