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Plant and Cell Physiology, 1992, Vol. 33, No. 2 165-170
© 1992

Synthesis of an S-(2-Aminoethyl)-L-Cysteine Conjugate in S-(2-Aminoethyl)-L-Cysteine- Resistant Adenine-Auxotrophic Cells of Datura innoxia Mill

Hitoshi Adachi, Masanori Aso1, Hiroyoshi Saito2 and Katsuhisa Furuhashi3

Department of Biology, Faculty of Science, Niigata University Niigata, 950-21 Japan

3To whom correspondence should be addressed

A line of S-(2-aminoethyl)-L-cysteine-resistant adenine-auxotrophic cells (Ad–AECr strain) was isolated from adenine-auxotrophic cells (Ad– strain) of Datura innoxia Mill by a stepwise selection method. Ad–AECr and Bl cells, which were cloned from the original Ad–AECr cells, were able to grow actively on medium that contained 10 mM S-(2-aminoethyl)-L-cysteine (AEC), whereas the growth of Ad– cells ceased completely in the presence of 0.5 mM AEC. The resistant phenotype has been maintained for at least 10 months in culture on medium without AEC.

Levels of free lysine in Ad–AECr and Bl cells were similar to that in Ad– cells. By contrast, the level of free AEC in Ad–AEC cells was 10-fold lower than in Ad– cells and no free AEC was detectable in Bl cells. However, acid hydrolysis of extracts from Ad–AECr and Bl cells resulted in a remarkable increase in levels of detectable AEC. This result indicates that conjugated AEC is synthesized and accumulated in the AEC-resistant cells. The level of the AEC conjugate in Bl cells increased with increases in the concentration of AEC in the culture medium, while intracellular levels of AEC were so low as not to be detectable in the case of cells grown on medium supplemented with AEC at less than 1 mM. The AEC conjugate was also detected in Ad– cells, but at lower levels than in the AEC-resistant cells. In addition, AEC was found to be incorporated into soluble proteins in Ad– cells.

These results suggest that the resistance of AEC-resistant cells of Datura innoxia is accomplished via acceleration of the synthesis of the AEC conjugate which prevents any increase in intracellular levels of free AEC.

1Present address: Institute for Biology and Chemistry, TsumuraCo. Ltd., Inashiki, Ibaraki, 300-03 Japan.

2Present address: North Kanto Shop, Sakata Seed Co. Ltd.,Saitama, 347 Japan.

(Received July 31, 1991; Accepted December 22, 1991)
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