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Sulfate reduction is increased in transgenic Arabidopsis thaliana expressing 5′‐adenylylsulfate reductase from Pseudomonas aeruginosa
Author(s) -
Tsakraklides George,
Martin Melinda,
Chalam Radhika,
Tarczynski Mitchell C.,
Schmidt Ahlert,
Leustek Thomas
Publication year - 2002
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1046/j.1365-313x.2002.01477.x
Subject(s) - sulfite reductase , thiosulfate , biochemistry , glutathione , sulfite , cysteine , arabidopsis thaliana , reductase , sulfurtransferase , biology , chemistry , glutathione reductase , sulfur metabolism , sulfite oxidase , glutaredoxin , enzyme , sulfur , mutant , gene , organic chemistry , glutathione peroxidase
Summary The two‐electron reduction of sulfate to sulfite in plants is mediated by 5′‐adenylylsulfate (APS) reductase, an enzyme theorized to be a control point for cysteine synthesis. The hypothesis was tested by expression in Arabidopsis thaliana under transcriptional control of the CaMV 35S promoter of the APS reductase from Pseudomonas aeruginosa ( Pa APR) fused with the rbcS transit peptide for localization of the protein to plastids. Pa APR was chosen for the experiment because it is a highly stable enzyme compared with the endogenous APS reductase of A. thaliana , and because Pa APR is catalytically active in combination with the plant thioredoxins m and f indicating that it would likely be catalytically active in plastids. The results indicate that sulfate reduction and O ‐acetylserine (OAS) production together limit cysteine synthesis. Transgenic A. thaliana lines expressing Pa APR accumulated sulfite, thiosulfate, cysteine, γ‐glutamylcysteine, and glutathione. Sulfite and thiosulfate increased more than did cysteine, γ‐glutamylcysteine and glutathione. Thiosulfate accumulation was most pronounced in flowers. Feeding of OAS to the Pa APR‐expressing plants caused cysteine and glutathione to increase more rapidly than in comparably treated wild type. Both wild‐type and transgenic plants accumulated sulfite and thiosulfate in response to OAS feeding. The Pa APR‐expressing plants were slightly chlorotic and stunted compared with wild type. An attempt to uncover the source of thiosulfate, which is not thought to be an intermediate of sulfate reduction, revealed that purified β‐mercaptopyruvate sulfurtransferase is able to form thiosulfate from sulfite and β‐mercaptopyruvate, suggesting that this class of enzymes could form thiosulfate in vivo in the presence of excess sulfite.