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Journal of Experimental Botany, Vol. 53, No. 378, pp. 2151-2158, November 1, 2002
© 2002 Oxford University Press

Recovery of tobacco cells from cadmium stress is accompanied by DNA repair and increased telomerase activity

Received 14 December 2001; Accepted 3 July 2002

Miloslava Fojtová1, Jana Fulnecková2, Jirí Fajkus1,2 and Ales Kovarík3,1

1 Institute of Biophysics, Academy of Sciences of the Czech Republic, Královopolská 135, Brno, CZ-612 65, Czech Republic
2 Department of Functional Genomics and Proteomics, Masaryk University Brno, CZ-612 65, Czech Republic

Abbreviations: dNTP, deoxynucleotide triphosphate; DTT, 1,4,-dithio-DL-threitol; FDA, fluorescein diacetate; Mb, megabase; MES, 2[N-morpholino]ethanesulphonic acid; MS medium, Murashige and Skoog medium; PFGE, pulsed-field-gel electrophoresis; Topoll, topoisomerase II; TRAP, telomere repeat amplifcation protocol.

It has been shown previously that apoptosis of tobacco cells induced by cadmium ions shows a relatively long lag period between exposure and cell death. This lag phase lasts for 3 d in TBY-2 cell cultures and is characterized by the maintenance of full cell viability despite extensive fragmentation of DNA into pieces of chromatin loop size. Experiments reported here demonstrate that cell death can be prevented if 50 µM CdSO4 is removed from the growth medium during the lag phase, suggesting that an irreversible apoptotic trigger is delivered within 24 h, between the third and fourth days of cadmium treatment. The post-cadmium recovery phase was characterized by DNA repair at the level of 50–200 kb and increased telomerase activity. Analysis of high-molecular-weight DNA by pulsed-field-gel electrophoresis revealed that the majority of DNA strand breaks was repaired within 48 h after cadmium withdrawal. Telomerase activity increased 2.5-fold in the recovery phase, but elevated levels were also found in cell extracts from apoptotic cells suggesting that telomerase might be associated with DNA repair, but it is not capable of inhibiting ongoing apoptosis. Limited exposure of TBY-2 cells to cadmium elicits non-random DNA damage of relatively high magnitude that can be repaired. It is proposed that plants might have developed a highly efficient DNA repair system to cope with transient genotoxic stress.

Key words: Key words: Apoptosis, cadmium, DNA domain fragmentation, telomerase, tobacco BY-2 cells.


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