Quantitative proteomics of heavy metal exposure in Arabidopsis thaliana reveals alterations in one-carbon metabolism enzymes upon exposure to zinc.
UNLABELLED
Plant zinc (Zn) homeostasis must be tightly regulated as the requirement for this micronutrient necessitates its uptake. However, excessive Zn can lead to toxicity and the plant must respond rapidly and effectively within its capacity to minimize damage. To detect mechanisms that may be important for coping with excess Zn we carried out a quantitative proteomics approach using 2D-DIGE to identify Zn-responsive proteins in microsomal fractions from leaves of 4day, 200μM Zn-treated, Arabidopsis thaliana plants. Of the eight proteins which showed significant changes in abundance in the Zn-treated samples and which met all of the selection criteria following statistical analysis, six were successfully identified by LC-MS/MS with 2 or more unique peptides. Three of the proteins were found to be involved in the one-carbon metabolism pathway; including glycine decarboxylase P protein, serine hydroxymethyltransferase (SHMT) and methionine synthase, all of which showed reduced abundance in the Zn-treated samples. Western blot analysis confirmed the decrease in SHMT, while changes in metal tolerance protein indicated plants were most likely actively sequestering Zn. Interestingly, excess Zn led to increased petiole length, a phenotype which may reflect the reduced levels of methionine, a key product of the one-carbon metabolism pathway.


BIOLOGICAL SIGNIFICANCE
Metal contamination is becoming an increasingly common environmental problem. High levels of zinc can be found in certain soils naturally or as a result of long-term anthropogenic activity which leads to its accumulation; i.e. use of fertilizers or industrial waste. The study of metal tolerant plants, particularly those classified as hyperaccumulators has been driven by the potential use of these plants for bioremediation purposes. However, the effects of heavy metal exposure on sensitive plants and the different cellular processes that are affected have received significantly less attention. We are interested in identifying proteins in A. thaliana that are induced as a result of exposure to subtoxic levels of heavy metals with the aim of discovering novel participants in heavy metal stress and adaptation.
['Barkla|Bronwyn J|BJ|', 'Vera-Estrella|Rosario|R|', 'Miranda-Vergara|María Cristina|MC|', 'Pantoja|Omar|O|']
D017360:Arabidopsis D029681:Arabidopsis Proteins D015153:Blotting, Western D002244:Carbon D002734:Chlorophyll D004396:Coloring Agents D015180:Electrophoresis, Gel, Two-Dimensional D012696:Glycine Hydroxymethyltransferase D006863:Hydrogen-Ion Concentration D008670:Metals D019216:Metals, Heavy D008715:Methionine D008861:Microsomes D018515:Plant Leaves D020543:Proteome D040901:Proteomics D012987:Soil D021241:Spectrometry, Mass, Electrospray Ionization D015032:Zinc
Micronutrients Abiotic stress DIGE Differential in-gel electrophoresis Mass spectrometry