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Related Experiment Videos

Nuclear DNA polymerases and the HeLa cell cycle.

R W Chiu, E F Baril

    The Journal of Biological Chemistry
    |October 10, 1975
    PubMed
    Summary

    HeLa cell DNA polymerase II activity rises during the cell cycle, suggesting its role in DNA replication. DNA polymerase I activity remains constant, indicating distinct cellular functions.

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    Area of Science:

    • Molecular Biology
    • Cell Biology
    • Biochemistry

    Background:

    • HeLa S3 cell nuclei possess two distinct DNA-dependent DNA polymerases.
    • Understanding the cell cycle variations in these enzyme activities is crucial for comprehending DNA replication regulation.

    Purpose of the Study:

    • To investigate the differential activities of two DNA polymerases during the HeLa cell cycle.
    • To elucidate the potential roles of DNA polymerase I and II in DNA biosynthesis.

    Main Methods:

    • Cell synchronization using a double thymidine block.
    • Partial purification of DNA polymerases via DEAE-cellulose and phosphocellulose chromatography.
    • Assaying enzyme activity at various cell cycle phases and in response to chemical inhibitors (cycloheximide, hydroxyurea).

    Main Results:

    • DNA polymerase I activity remained constant throughout the cell cycle.
    • DNA polymerase II activity significantly increased from G1 to mid-S phase and then decreased sharply by G2.
    • Cycloheximide abolished the rise in DNA polymerase II activity, while hydroxyurea had no effect.

    Conclusions:

    • DNA polymerase I and II are distinct enzymes with differing roles in HeLa cells.
    • DNA polymerase II likely functions in DNA replication and its activity is regulated by protein synthesis and cell cycle progression.
    • The findings suggest DNA polymerase II is induced during periods of DNA biosynthesis.

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