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Related Experiment Videos

A ribo-deoxyribonucleotide primer synthesized by primase.

L Rowen, A Kornberg

    The Journal of Biological Chemistry
    |February 10, 1978
    PubMed
    Summary

    Primase synthesizes shorter RNA primers for phage G4 DNA replication in the presence of deoxynucleoside triphosphates (dNTPs). These shorter primers remain effective for DNA polymerase III holoenzyme replication.

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    Area of Science:

    • Molecular Biology
    • Virology
    • Biochemistry

    Background:

    • Phage G4 DNA replication initiates with a ribonucleotide primer synthesized by primase.
    • The precise length and composition of this primer influence replication efficiency.

    Purpose of the Study:

    • To investigate the effect of deoxynucleoside triphosphates (dNTPs) on primase-synthesized primer length.
    • To determine the functional impact of altered primers on DNA replication by DNA polymerase III holoenzyme.

    Main Methods:

    • Enzymatic synthesis of RNA primers by primase in the presence of varying concentrations of ribonucleoside triphosphates (rNTPs) and dNTPs.
    • Analysis of primer length and composition.
    • Assay of primer efficiency in initiating DNA replication using DNA polymerase III holoenzyme.

    Main Results:

    • Primase synthesized shorter RNA primers (≤6 residues) when dNTPs were present at 50 μM with rNTPs at 20 μM.
    • These shorter primers were still effective in priming replication by DNA polymerase III holoenzyme.
    • Guanosine triphosphate (GTP) stimulated DNA replication, suggesting its incorporation enhances primer efficiency.

    Conclusions:

    • Primase can incorporate dNTPs into primers, forming hybrid ribonucleotide-deoxyribonucleotide chains.
    • While dNTP incorporation is less efficient than rNTPs, it allows for primer modification.
    • The presence of all four dNTPs significantly inhibits RNA synthesis by primase.

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