Identification of glutathione-S-transferase m19 and m34 among responsive GST genes against 1-chloro-2,4-dinitrobenzene treatment of Tetrahymena thermophila.

Industrial xenobiotic pollutants have toxic effects on diverse organisms in their natural environments. This study aims to identify the Glutathione-S-transferases (GST) from Tetrahymena thermophila that are highly responsive to the treatment of synthetic substrate 1-chloro-2,4-dinitrobenzene (CDNB). The LD50 value of CDNB was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test as 0.079 mM at 9 h exposure. The glutathione affinity-purified 22 kDa and 23 kDa GSTs from CDNB-treated cells were identified as GSTm19 and GSTm34 with 2D-gel electrophoresis coupled MALDI-Tof MS/MS analysis. The specific activitiy of the affinity-purified GSTs was upregulated upon the treatment of 0.072 mM CDNB with the decreased cell survival. GSTm19 and GSTm34 had also upregulated the mRNA expression under the highest dose treatment. The high cell survival and elevated total GST enzyme activity at 9 h under CDNB doses could be the result of both transcriptional upregulations as well as post-translational modifications. As a result, the cell survival of Tetrahymena thermophila was significantly affected by CDNB exposure in a concentration-dependent manner with the effect of low-dose stimulation and high-dose inhibition.