ABSTRACT
γ-Glutamyl transferases (GGT; EC 2.3.2.2) are glutathione-degrading enzymes that are represented in Arabidopsis thaliana by a small gene family of four members. Two isoforms, GGT1 and GGT2, are apoplastic, sharing broad similarities in their amino acid sequences, but they are differently expressed in the tissues: GGT1 is expressed in roots, leaves, and siliques, while GGT2 was thought to be expressed only in siliques. It is demonstrated here that GGT2 is also expressed in wild-type roots, albeit in very small amounts. GGT2 expression is enhanced in ggt1 knockout mutants, suggesting a compensatory effect to restore GGT activity in the root apoplast. Supplementation with 100 µM glutathione (GSH) resulted in the up-regulation of GGT2 gene expression in wild-type and ggt1 knockout roots, and of GGT1 gene expression in wild-type roots. Glutathione recovery was hampered by the GGT inhibitor serine/borate, suggesting a major role for apoplastic GGTs in this process. These findings can explain the ability of ggt1 knockout mutants to retrieve exogenously added glutathione from the growth medium.
Subject(s)
Arabidopsis/enzymology , Gene Expression Regulation, Enzymologic , Glutathione/metabolism , gamma-Glutamyltransferase/metabolism , Arabidopsis/genetics , Gene Expression Regulation, Plant , Plant Leaves/enzymology , Plant Leaves/genetics , Plant Leaves/metabolism , Plant Roots/enzymology , Plant Roots/genetics , Plant Roots/metabolism , gamma-Glutamyltransferase/chemistry , gamma-Glutamyltransferase/geneticsABSTRACT
Gamma-glutamyl transferase/transpeptidase (GGT, (5-l-glutamyl)-peptide:amino-acid 5-glutamyltransferase; EC 2.3.2.2.) is an ectoenzyme promoting the cleavage of the gamma-glutamyl moiety of glutathione (GSH) and gamma-glutamyl related compounds. In this work, we describe the localization of GGT by enzymehistochemical and immunohistochemical analysis in maize plants. Our results show that the tissue spatial distribution of GGT activity closely correlates with the localization of the GGT protein. We also demonstrate that GGT activity and protein are unevenly distributed in tissues, being higher in the epidermis and stomata, parenchyma of conductive elements and root meristem. These results can contribute to our understanding of GGT function and regulation as well as its role in glutathione metabolism. To date, these are largely unknown in plants.
Subject(s)
Gene Expression Regulation, Plant , Zea mays/enzymology , gamma-Glutamyltransferase/metabolism , Electrophoresis, Polyacrylamide Gel , Immunoblotting , Organ Specificity , Plant Leaves/enzymology , Plant Roots/cytology , Plant Roots/enzymology , Plant Shoots/cytology , Plant Shoots/enzymology , Zea mays/genetics , gamma-Glutamyltransferase/geneticsABSTRACT
Two maize genotypes differently responsive to nitrogen availability were characterized for their efficiency in nitrate accumulation via both the LATS (Low-Affinity Transport System) and HATS (High-Affinity Transport System) nitrate uptake systems. In addition, a full-length cDNA encoding a putative high-affinity nitrate transporter (ZmNrt2.1) was isolated and its expression evaluated in both the roots and leaves of the two maize genotypes, together with the expression of a maize H(+)-ATPase isoform (Mha1). The data showed the importance of the iHATS (Inducible High-Affinity System) system efficiency as a physiological marker of adaptation to low input and suggested that the transcript accumulation of ZmNrt2.1 might be a key step for the regulation of iHATS. However, ZmNrt2.1 transcription cannot account for the differences found between the two hybrids in terms of the activity of their respective iHATS and, as a consequence, of their adaptation to low input. Therefore, the involvement of some other transporter(s) or of some post-transcriptional/post-translational mechanism of regulation affecting the efficiency of iHATS may be hypothesized. In addition, the data suggest that the transcription of the Mha1 gene may also be involved in the global efficiency of the iHATS system.
