Blockade of Telomerase Function by Nucleoside Analogs

Y. E. Yegorov,^1,2 D. N. Chernov,^1,3 S. S. Akimov,¹ A. K. Akhmalisheva,⁴ Y. B. Smirnova,¹ D. B. Shinkarev,¹ I. V. Semenova,⁴ I. N. Yegorova,¹ and A. V. Zelenin¹

¹Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, ul. Vavilova 32, Moscow, 117984 Russia; fax: (095) 135-1405; E-mail: yegorov@genome.eimb.rssi.ru

²To whom correspondence should be addressed.

³Center of Medical Studies of Oslo University, ul. Vavilova 32, Moscow, 117984 Russia.

⁴Department of Embryology, School of Biology, Lomonosov Moscow State University, 119899 Russia.

Submitted June 30, 1997.

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Two types of spontaneously transformed cells appear in the culture of senescent mouse embryonic fibroblasts. The first type are cells with restricted proliferative potential (up to 30 population doublings); the other type are immortalized cells. Cells of the first type, unlike those of the second, have no telomerase activity and undergo two rounds of senescence. Spontaneous transformation of mouse embryonic fibroblasts in the presence of the reverse transcriptase inhibitors azidothymidine and carbovir led to the formation of telomerase-free clones. A fraction of these clones have the ability to overcome senescence via the acquisition of high telomerase activity. Cells with a very high level of telomerase activity become resistant to azidothymidine and carbovir. Azidothymidine-induced artificial senescence of rat myoblasts in culture resembles the senescence of fibroblasts, but the resulting cells acquire sharp morphological peculiarities. The blockade of telomerase function by azidothymidine in human U-937 and MeWo cells leads to the shortening of telomeres, but does not result in senescence. A hypothesis of the generation of the signal that induces senescence is proposed. This hypothesis suggests a change in DNA conformation during telomere shortening as a result of a change of loop structure of telomeric chromatin.

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KEY WORDS: telomerase, senescence, reverse transcriptase inhibitors, cell culture, differentiation, telomeres, spontaneous transformation.

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