Molecular cloning of AbGst1 encoding a glutathione transferase differentially expressed during exposure of Alternaria brassicicola to isothiocyanates.
The AbGst1 gene encoding a glutathione transferase from the necrotrophic pathogen Alternaria brassicicola was cloned from a benzyl isothiocyanate-treated conidial culture using differential display reverse transcription. The deduced amino-acid sequence of AbGst1p showed a significant degree of similarity to glutathione transferase-I from Saccharomyces cerevisiae and glutathione transferase-III from Schizosaccharomyces pombe. The transcription of AbGst1 was significantly enhanced by isothiocyanates, heavy metals and 1-chloro-2,4-dinitrobenzene. However, no significant transcript response was obtained with superoxide-generating menadione and paraquat. Recombinant AbGst1p expressed in Escherichia coli exhibited high transferase activity with allyl and benzyl isothiocyanates as substrate compared with 1-chloro-2,4-dinitrobenzene, but no peroxidase activity was detected. AbGst1 was upregulated in planta during the first day postinfection, suggesting the potential involvement of this enzyme in isothiocyanate detoxification mechanisms during host plant infection.
Sellam
Adnane
A
UMR PAVE Faculté des Sciences, Angers, France.
Poupard
Pascal
P
Simoneau
Philippe
P
D000528:Alternaria / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000382:microbiology; D001483:Base Sequence; D003001:Cloning, Molecular; D004137:Dinitrochlorobenzene / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D005982:Glutathione Transferase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D017879:Isothiocyanates / Q000494:pharmacology*; D019216:Metals, Heavy / Q000494:pharmacology; D008969:Molecular Sequence Data; D010802:Phylogeny; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid; D015854:Up-Regulation
D004137:Dinitrochlorobenzene; D005656:Fungal Proteins; D017879:Isothiocyanates; D019216:Metals, Heavy; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase