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Genome-wide Determination of Mammalian Replication Timing by DNA Content Measurement
08:06

Genome-wide Determination of Mammalian Replication Timing by DNA Content Measurement

Published on: January 19, 2017

[Increased expression of genes coding for replication proteins during the period of DNA synthesis]

O P Zhuchenko, L T Timchenko, N A Timchenko

    Molekuliarnaia Biologiia
    |May 1, 1990
    PubMed

    Insights

    Researchers identified key proteins in the alpha-polymerase complex from liver cells. One protein (pr167) shows DNA polymerase activity, while another (pr12) may initiate DNA replication.

    Area of Science:

    • Molecular Biology
    • Hepatocyte Research
    • DNA Replication

    Background:

    • The alpha-polymerase complex is crucial for DNA replication.
    • Understanding its components is essential for studying cell proliferation and liver regeneration.

    Purpose of the Study:

    • To isolate and characterize individual messenger RNAs (mRNAs) encoding proteins of the alpha-polymerase complex.
    • To identify novel components involved in DNA replication initiation.

    Main Methods:

    • Isolation of mRNAs from replicating hepatocytes.
    • Synthesis of complementary DNAs (cDNAs) and identification of clones via colony hybridization.
    • Hybrid-selection of specific mRNAs and cell-free translation.
    • Analysis of translation products using activity gel electrophoresis.

    Main Results:

    • Three cDNA clones (pr12, pr127, pr167) complementary to alpha-polymerase mRNAs were identified.
    • pr167 mRNA codes for a 140 kDa protein with DNA polymerase activity.
    • pr12 mRNA codes for a protein, potentially a novel component of the alpha-polymerase complex.

    Conclusions:

    • pr167 is a DNA polymerase subunit within the alpha-polymerase complex.
    • pr12 is a newly identified protein that may play a role in initiating DNA replication.
    • These findings contribute to understanding the molecular mechanisms of DNA replication and liver regeneration.

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