Enzyme activities and subcellular localization of members of the Arabidopsis glutathione transferase superfamily.
Enzyme screens with Strep-tagged recombinant proteins and expression studies with the respective green fluorescent protein (GFP) fusions have been employed to examine the functional activities and subcellular localization of members of the Arabidopsis glutathione transferase (GST) superfamily. Fifty-one of 54 GST family members were transcribed and 41 found to express as functional glutathione-dependent enzymes in Escherichia coli. Functional redundancy was observed and in particular three theta (T) class GSTs showed conserved activities as hydroperoxide-reducing glutathione peroxidases (GPOXs). When expressed in tobacco as GFP fusions, all three GSTTs localized to the peroxisome, where their GPOX activity could prevent membrane damage arising from fatty acid oxidation. Through alternative splicing, two of these GSTTs form fusions with Myb transcription factor-like domains. Examination of one of these variants showed discrete localization within the nucleus, possibly serving a role in reducing nucleic acid hydroperoxides or in signalling. Based on this unexpected differential sub-cellular localization, 15 other GST family members were expressed as GFP fusions in tobacco. Most accumulated in the cytosol, but GSTU12 localized to the nucleus, a family member resembling a bacterial tetrachlorohydroquinone dehalogenase selectively associated with the plasma membrane, and a lambda GSTL2 was partially directed to the peroxisome after removal of a putative chloroplast transit peptide. Based on the results obtained with the GSTTs, it was concluded that these proteins can exert identical protective functions in differing subcellular compartments.
Dixon
David P
DP
School of Biological and Biomedical Sciences, Durham University, Durham DH1 3LE, UK.
Hawkins
Timothy
T
Hussey
Patrick J
PJ
Edwards
Robert
R
D020816:Amino Acid Motifs; D017360:Arabidopsis / Q000201:enzymology*; D003600:Cytosol / Q000201:enzymology; D005822:Genetic Vectors / Q000235:genetics; D005982:Glutathione Transferase / Q000737:chemistry / Q000378:metabolism*; D042541:Intracellular Space / Q000201:enzymology; D018613:Microscopy, Confocal; D010802:Phylogeny; D021381:Protein Transport; D011993:Recombinant Fusion Proteins; D013347:Subcellular Fractions / Q000201:enzymology; D014026:Nicotiana / Q000166:cytology / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase