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Deoxyuridine triphosphate pools after polyoma virus infection
The Journal of Biological Chemistry
|October 25, 1980
Summary
Polyoma DNA synthesis in mouse cells produces short fragments. Deoxyuridine triphosphate (dUTP) incorporation minimally contributes to these fragments, even when DNA synthesis is inhibited by hydroxyurea.
Area of Science:
- Molecular Biology
- Virology
- Biochemistry
Background:
- Polyoma DNA synthesis is discontinuous, forming Okazaki fragments.
- Hydroxyurea inhibits DNA synthesis by targeting ribonucleotide reductase.
- Inhibition leads to increased incorporation into short DNA fragments.
Purpose of the Study:
- To investigate the role of deoxyuridine triphosphate (dUTP) incorporation in short fragment formation during polyoma DNA synthesis.
- To quantify dUTP levels in infected cells and assess its contribution to DNA repair.
Main Methods:
- Development of a sensitive enzymatic assay to measure dUTP pools relative to deoxythymidine triphosphate (dTTP).
- Analysis of dUTP levels in mouse fibroblasts before and after polyoma virus infection.
- Assessment of dUTP levels under hydroxyurea-induced inhibition of DNA synthesis.
Main Results:
- No detectable dUTP was found in growing mouse fibroblasts.
- Infected cells showed 0.4% dUTP (of dTTP) at peak DNA synthesis.
- Hydroxyurea treatment caused dUTP levels to decrease.
Conclusions:
- Deoxyuridine triphosphate incorporation plays a minimal role in generating short DNA fragments during polyoma DNA synthesis.
- The observed short fragments are unlikely to be primarily due to dUTP incorporation and subsequent repair.