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Sequence-dependent DNA replication in preimplantation mouse embryos.

D O Wirak, L E Chalifour, P M Wassarman

    Molecular and Cellular Biology
    |November 1, 1985
    PubMed
    Summary
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    Genome Duplication at the Beginning of Mammalian Development.

    Current topics in developmental biology·2016

    Specific DNA sequences, like polyomavirus (PyV) or simian virus 40 (SV40), are essential for DNA replication in mouse embryos. This replication is sequence-dependent and requires specific viral proteins for activation.

    Area of Science:

    • Molecular Biology
    • Developmental Biology
    • Virology

    Background:

    • Investigating DNA replication mechanisms is crucial for understanding embryonic development.
    • Previous studies suggested sequence-independent DNA replication in some early developmental stages.

    Purpose of the Study:

    • To determine if specific DNA sequences are required for DNA replication during mouse embryonic development.
    • To identify the viral elements necessary for DNA replication in early mouse embryos.

    Main Methods:

    • Injection of circular, double-stranded DNA molecules (plasmid pML-1, polyomavirus (PyV) DNA, simian virus 40 (SV40) DNA) into mouse oocytes and one- or two-cell embryos.
    • Analysis of DNA stability and replication using molecular techniques.
    • Investigating the role of PyV large T-antigen and origin of replication configurations.

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    Main Results:

    • Plasmid pML-1 DNA replicated only when containing PyV or SV40 sequences.
    • Replication occurred in embryos but not oocytes, indicating developmental stage-specific permissiveness.
    • PyV DNA replicated efficiently, requiring PyV large T-antigen and specific origin configurations, demonstrating cell-specific activation.
    • SV40 DNA replicated less efficiently than PyV DNA.

    Conclusions:

    • Mammalian embryonic DNA replication requires specific cis-acting sequences, unlike Xenopus egg replication.
    • Mouse embryos are permissive for PyV DNA replication, dependent on viral factors and specific origin sequences.
    • Replication of viral DNA can interfere with normal embryonic development.