Cloning and molecular analysis of radish (Raphanus sativus L.) cDNAs encoding heterodimeric .GAMMA.-glutamyltransferases

Nakano, Yuzo; Prieto, Rafael; Hara, Masakazu; Sekiya, Jiro

doi:10.5511/plantbiotechnology.23.419

Cited by 2 publications

(5 citation statements)

References 21 publications

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“…Based on the phylogenetic analysis, VvGGT3 groups with Arabidopsis GGT4 (AtGGT4) and garlic GGT2 (AsGGT2) both of which have been functionally shown to be located in the vacuole (Grzam et al, 2007; Yoshimoto et al, 2015). Additionally, radish has three at least GGT enzymes (Nakano et al, 2006b) of which RsGGT3 is the closest match to VvGGT3 ( Figure 1 ). When over-expressed in tobacco, RsGGT3 unlike RsGGT1 and RsGGT2 which are most likely bound to cell walls was able to be purified from the soluble fraction (Nakano et al, 2006b), suggesting that it too like AtGGT4 may be localized to the vacuole (Ohkama-Ohtsu et al, 2009).…”

Section: Resultsmentioning

confidence: 99%

“…Additionally, radish has three at least GGT enzymes (Nakano et al, 2006b) of which RsGGT3 is the closest match to VvGGT3 ( Figure 1 ). When over-expressed in tobacco, RsGGT3 unlike RsGGT1 and RsGGT2 which are most likely bound to cell walls was able to be purified from the soluble fraction (Nakano et al, 2006b), suggesting that it too like AtGGT4 may be localized to the vacuole (Ohkama-Ohtsu et al, 2009). Removal of the putative N-terminal leader/targeting peptides and reconstruction of the phylogenetic tree does not substantially affect the sequence relationships observed ( Supplementary Figure 1 ).…”

Section: Resultsmentioning

confidence: 99%

“…In plants, there has been comparatively little research focus on GGTs. However, GGTs from a range of model as well as non-model plant species have begun to be functionally characterized (Nakano et al, 2006b; Yoshimoto et al, 2015). In Arabidopsis thaliana ( Arabidopsis ), where plant GGTs have been most extensively studied, four GGT genes have been identified, of which three are functional ( AtGGT1 , AtGGT2 , and AtGGT4 ) (Grzam et al, 2007; Martin et al, 2007; Ohkama-Ohtsu et al, 2007a; Tolin et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

“…There are three functional GGT genes in radish, RsGGT1 (BAC45233.1), RsGGT2 (BAC56855.1), and RsGGT3 (BAD22536.1). These genes have been used to assist in the classification of plant GGTs, which fall into two groups based on their solubility (either soluble or bound) (Nakano et al, 2006a; Nakano et al, 2006b). In addition, they have been used to classify GGTs based molecular structures, these include the monomeric GGT and heterodimeric GGT, both of which present different substrate specificities (Nakano et al, 2006a; Nakano et al, 2006b).…”

Section: Introductionmentioning

confidence: 99%

“…These genes have been used to assist in the classification of plant GGTs, which fall into two groups based on their solubility (either soluble or bound) (Nakano et al, 2006a; Nakano et al, 2006b). In addition, they have been used to classify GGTs based molecular structures, these include the monomeric GGT and heterodimeric GGT, both of which present different substrate specificities (Nakano et al, 2006a; Nakano et al, 2006b). Similarly, three functional GGT genes are present in garlic, AsGGT1 (BAQ21911.1), AsGGT2 (BAQ21912.1), and AsGGT3 (BAQ21913.1) (Yoshimoto et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

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Functional Characterization of the Grapevine γ-Glutamyl Transferase/Transpeptidase (E.C. 2.3.2.2) Gene Family Reveals a Single Functional Gene Whose Encoded Protein Product Is Not Located in Either the Vacuole or Apoplast

2019

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γ-glutamyl transferases/transpeptidases (E.C. 2.3.2.2, GGTs) are involved in the catabolism of many compounds that are conjugated to glutathione (GSH), which have a variety of roles. GSH can act as storage and transport vehicle for reduced sulfur; it is involved in the detoxification of xenobiotics and also acts as a redox buffer by utilizing its thiol residue to protect against reactive oxygen species, which accumulate in response to biotic and abiotic stress. Furthermore, many distinctive flavor and aroma compounds in Sauvignon blanc wines originate from odorless C5- and C6-GSH conjugates or their GGT catabolized derivatives. These precursors are then processed into their volatile forms by yeast during fermentation. In many plant species, two or more isoforms of GGTs exist that target GSH-conjugates to either the apoplast or the vacuole. A bioinformatics approach identified multiple GGT candidates in grapevine (Vitis vinifera). However, only a single candidate, VvGGT3, has all the conserved residues needed for GGT activity. This is intriguing given the variety of roles of GSH and GGTs in plant cells. Characterization of VvGGT3 from cv. Sauvignon blanc was then undertaken. The VvGGT3 transcript is present in roots, leaves, inflorescences, and tendril and at equal abundance in the skin, pulp, and seed of mature berries and shows steady accumulation over the course of whole berry development. In addition, the VvGGT3 transcript in whole berries is upregulated upon Botrytis cinerea infection as well as mechanical damage to leaf tissue. VvGGT3-GFP fusion proteins transiently over-expressed in onion cells were used to study subcellular localization. To confirm VvGGT3 activity and localization in vivo, the fluorescent γ-glutamyl-7-amido-4-methylcoumarin substrate was added to Nicotiana benthamiana leaves transiently over-expressing VvGGT3. In combination, these results suggest that the functional VvGGT3 is associated with membrane-like structures. This is not consistent with its closely related functionally characterized GGTs from Arabidopsis, radish and garlic.

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