Polyamines as Modulators of Gene Expression under Oxidative Stress in Escherichia coli

A. G. Tkachenko^* and L. Yu. Nesterova

Institute of Ecology and Genetics of Microoragnisms, Ural Division of the Russian Academy of Sciences, ul. Goleva 13, Perm 614081, Russia; fax: (3422) 646-711; E-mail: agtkachenko@ecology.psu.ru

^* To whom correspondence should be addressed.

Received April 1, 2002; Revision received July 2, 2002

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Activity of enzymes of polyamine synthesis and contents of their products increased in E. coli cells in response to oxidative stress caused by addition of hydrogen peroxide to an exponentially growing culture. Putrescine and spermidine added to the culture medium in physiological concentrations significantly increased expression of genes oxyR and katG responsible for defense against oxidative stress, whereas cadaverine had no effect. The role of polyamines as modulators of the gene expression was confirmed by experiments with an inhibitor of polyamine synthesis, 1,3-diaminopropane, which decreased the level of cell polyamines and thus abolished the ability of the cell to induce oxyR expression under oxidative stress. A genetic method gave similar results: under oxidative stress mutants with disorders in polyamine synthesis displayed a significantly decreased level of induction of the oxyR and katG genes, and this level was recovered on addition of putrescine. In the presence of inhibitors of DNA-gyrase, nalidixic acid and novobiocin, the oxyR expression depended on the extent of DNA supercoiling. Putrescine decreased the inhibitory effects of nalidixic acid and novobiocin, and this confirmed its properties of a stimulator of DNA supercoiling. Resistance to rifampicin was studied to exemplify the mutation rate under oxidative stress. Putrescine decreased twofold the level of mutations and increased the number of viable cells in the culture exposed to oxidative stress.

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KEY WORDS: polyamines, oxidative stress, oxyR, modulators, gene expression, DNA topology, survival, mutation frequency

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