Effect of Human Cell Malignancy on Activity of DNA Polymerase ι

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A. A. Kazakov¹, E. E. Grishina², V. Z. Tarantul^1*, and L. V. Gening¹

¹Institute of Molecular Genetics, Russian Academy of Sciences, pl. Kurchatova 2, 123182 Moscow, Russia; fax: (499) 196-0221; E-mail: tarantul@img.ras.ru

²Ophthalmological Clinical Hospital, Moscow Department of Health, Mamonovsky Pereulok 7, 103001 Moscow, Russia; fax: (495) 650-1822; E-mail: eyelena@mail.ru

^* To whom correspondence should be addressed.

Received December 30, 2009; Revision received February 19, 2010

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An increased level of mutagenesis, partially caused by imbalanced activities of error prone DNA polymerases, is a key symptom of cell malignancy. To clarify the possible role of incorrect DNA polymerase ι (Pol ι) function in increased frequency of mutations in mammalian cells, the activity of this enzyme in extracts of cells of different mouse organs and human eye (melanoma) and eyelid (basal-cell skin carcinoma) tumor cells was studied. Both Mg²⁺, considered as the main activator of the enzyme reaction of in vivo DNA replication, and Mn²⁺, that activates homogeneous Pol ι preparations in experiments in vitro more efficiently compared to all other bivalent cations, were used as cofactors of the DNA polymerase reaction in these experiments. In the presence of Mg²⁺, the enzyme was active only in cell extracts of mouse testicles and brain, whereas in the presence of Mn²⁺ the activity of Pol ι was found in all studied normal mouse organs. It was found that in cell extracts of both types of malignant tumors (basal-cell carcinoma and melanoma) Pol ι activity was observed in the presence of either Mn²⁺ or Mg²⁺. Manganese ions activated Pol ι in both cases, though to a different extent. In the presence of Mn²⁺ the Pol ι activity in the basal-cell carcinoma exceeded 2.5-fold that in control cells (benign tumors from the same eyelid region). In extracts of melanoma cells in the presence of either cation, the level of the enzyme activity was approximately equal to that in extracts of cells of surrounding tumor-free tissues as well as in eyes removed after traumas. The distinctive feature of tissue malignancy (in basal-cell carcinoma and in melanoma) was the change in DNA synthesis revealed as Mn²⁺-activated continuation of DNA synthesis after incorrect incorporation of dG opposite dT in the template by Pol ι. Among cell extracts of different normal mouse organs, only those of testicles exhibited a similar feature. This similarity can be explained by cell division blocking that occurs in all normal cells except in testicles and in malignant cells.

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KEY WORDS: uveal eye melanoma, basal-cell carcinoma, DNA polymerase ι, TLS

DOI: 10.1134/S0006297910070138

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