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Proofreading and DNA Repair Assay Using Single Nucleotide Extension and MALDI-TOF Mass Spectrometry Analysis
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Proofreading and DNA Repair Assay Using Single Nucleotide Extension and MALDI-TOF Mass Spectrometry Analysis

Published on: June 19, 2018

[Effect of Mn(II) on the error-prone DNA polymerase iota activity in extracts from human normal and tumor cells]

A V Lakhin, A S Efremova, I V Makarova

    Molekuliarnaia Genetika, Mikrobiologiia I Virusologiia
    |June 22, 2013
    PubMed

    Insights

    Manganese (Mn2+) activates error-prone DNA polymerase iota (Pol iota) in normal and tumor cells. An RNA aptamer (IKL5) effectively suppressed this enhanced Pol iota activity in tumor cells, suggesting therapeutic potential.

    Area of Science:

    • Biochemistry
    • Molecular Biology
    • Genetics

    Context:

    • DNA polymerase iota (Pol iota) exhibits unique, error-prone DNA synthesis.
    • Pol iota is preferentially activated by manganese ions (Mn2+) over magnesium ions (Mg2+).
    • Understanding Mn2+-dependent DNA synthesis is crucial for cancer research.

    Purpose:

    • To investigate the effect of Mn2+ on DNA synthesis in various human and murine cell extracts.
    • To characterize Mn2+-dependent DNA synthesis in normal tissues, uveal melanoma, and cancer cell lines (SKOV-3, HL-60).
    • To evaluate the efficacy of an RNA aptamer (IKL5) in suppressing Pol iota activity.

    Summary:

    • Mn2+ significantly influences DNA synthesis across normal and malignant cell types, with distinct patterns observed.
    • Malignant transformation alters Mn2+-dependent DNA synthesis.
    • The RNA aptamer IKL5 demonstrated efficient suppression of error-prone DNA synthesis by Pol iota in all tested cell extracts.

    Impact:

    • The study reveals characteristic Mn2+-dependent DNA synthesis profiles in normal and tumor cells.
    • Identifies changes associated with malignant transformation.
    • Suggests IKL5 as a potential therapeutic agent to inhibit enhanced Pol iota activity in cancer cells.

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