The Reparative dna Polymerase eta (Polη) Plays a key Role in Mutagenesis at Low Doses of uv Radiation in Yeast Saccharomyces cerevisiae

Мұқаба

Дәйексөз келтіру

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Ашық рұқсат Ашық рұқсат
Рұқсат жабық Рұқсат берілді
Рұқсат жабық Тек жазылушылар үшін

Аннотация

Under normal conditions and when exposed to low doses of mutagens, DNA damage tolerance systems (DDT) are a key way to combat DNA damage in bacterial and eukaryotic cells. Two different DDT pathways, erroneous (translesion synthesis – TLS) and error-free (recombination), operate in all eukaryotic organisms. TLS involves the protein complex polymerase zeta (Polζ) (encoded by the genes REV1, REV3 and REV7), and polymerase eta (Polη) (encoded by the RAD30 gene); these polymerases are conserved from yeast to humans. We have shown that at low levels of UV radiation doses, a decisive role in mutagenesis is played by the high activity of Poly, which bypasses the overwhelming number of DNA damage that got in the way of the replication machine. Both in the case of irradiation with high and low doses of UV rays, checkpoint plays an important role in induced mutagenesis. However, the roles of genes involved in the regulation of UV-induced mutagenesis at low and high doses often differ. Inactivation of the RAD30 gene at low doses leads to an increased level of induced mutagenesis, and at high doses it practically does not differ from the level of the wild-type strain. Deletions of the HIM1, HSM3 and HIF1 genes, on the contrary, reduce the high level of mutagenesis characteristic of high doses to the level of the wild-type strain at low doses. These differences characterize the local and global checkpoint.

Толық мәтін

Рұқсат жабық

Авторлар туралы

E. Alekseeva

Konstantinov Petersburg Nuclear Physics Institute of the National Research Centre “Kurchatov Institute”; Kurchatov Genome Center — Petersburg Nuclear Physics Institute

Хат алмасуға жауапты Автор.
Email: alekseeva_ea@pnpi.nrcki.ru
Ресей, Leningrad oblast, Gatchina, 188300; Leningrad oblast, Gatchina, 188300

T. Evstyukhina

Konstantinov Petersburg Nuclear Physics Institute of the National Research Centre “Kurchatov Institute”; Kurchatov Genome Center — Petersburg Nuclear Physics Institute

Email: alekseeva_ea@pnpi.nrcki.ru
Ресей, Leningrad oblast, Gatchina, 188300; Leningrad oblast, Gatchina, 188300

I. Skobeleva

Konstantinov Petersburg Nuclear Physics Institute of the National Research Centre “Kurchatov Institute”

Email: alekseeva_ea@pnpi.nrcki.ru
Ресей, Leningrad oblast, Gatchina, 188300

V. Peshekhonov

Konstantinov Petersburg Nuclear Physics Institute of the National Research Centre “Kurchatov Institute”; Kurchatov Genome Center — Petersburg Nuclear Physics Institute

Email: alekseeva_ea@pnpi.nrcki.ru
Ресей, Leningrad oblast, Gatchina, 188300; Leningrad oblast, Gatchina, 188300

V. Korolev

Konstantinov Petersburg Nuclear Physics Institute of the National Research Centre “Kurchatov Institute”; Kurchatov Genome Center — Petersburg Nuclear Physics Institute

Email: alekseeva_ea@pnpi.nrcki.ru
Ресей, Leningrad oblast, Gatchina, 188300; Leningrad oblast, Gatchina, 188300

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2. Fig. 1. Sensitivity to UV light and the frequency of UV-induced mutagenesis at the CAN1 locus in the wild-type strain and the rad30Δ, pms1Δ and pms1Δ rad30Δ mutant strains treated with 7, 14 and 21 J/m² of UV radiation. The graphs show the standard errors of the mean (± SEM) obtained from five independent experiments.

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3. Fig. 2. Sensitivity to UV light and the frequency of UV-induced mutagenesis at the CAN1 locus in the wild-type strain and the rad30Δ, him1Δ and him1Δ rad30Δ mutant strains when treated with doses of 7, 14 and 21 J/m² UV irradiation. The graphs show standard errors of the mean (± SEM) obtained from five independent experiments.

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4. Fig. 3. Relative normalized expression of the RNR3 gene in the wild-type strain and the him1Δ and hif1Δ mutant strains before and after irradiation with ultraviolet light (after UV irradiation, the cells were kept for four hours at 30 °C in an induction thermostat); the UV dose was 14 J/m²; * p < 0.05, Student's t-test.

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5. Fig. 4. Sensitivity to UV light and the frequency of UV-induced mutagenesis at the CAN1 locus in the wild-type strain and the rad30Δ, hsm3Δ, hif1Δ, hsm3Δ rad30Δ and hif1Δ rad30Δ mutant strains treated with 7, 14 and 21 J/m² of UV irradiation. The graphs show the standard errors of the mean (± SEM) obtained from five independent experiments.

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6. Fig. 5. Sensitivity to UV light and the frequency of UV-induced mutagenesis at the CAN1 locus in the wild-type strain and the rad30Δ, sml1Δ, rad53 + HA-F, sml1Δ rad30Δ, and rad53 + HA-F rad30Δ mutant strains treated with 7, 14, and 21 J/m² of UV irradiation. The graphs show the standard errors of the mean (±SEM) obtained from five independent experiments.

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7. Fig. 6. Sensitivity to UV light and the frequency of UV-induced mutagenesis at the CAN1 locus in the wild-type strain and the hat1Δ mutant strain when treated with doses of 7, 14 and 21 J/m² of UV irradiation. The graphs show standard errors of the mean (± SEM) obtained from five independent experiments.

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