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Related Experiment Videos

Polyomavirus enhancer contains multiple redundant sequence elements that activate both DNA replication and gene

G M Veldman, S Lupton, R Kamen

    Molecular and Cellular Biology
    |April 1, 1985
    PubMed
    Summary

    Polyomavirus enhancer sequences are crucial for viral DNA replication and transcriptional enhancement. This study identified four functionally redundant elements within the enhancer region essential for activating replication, with multiple copies restoring function.

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    Area of Science:

    • Molecular Biology
    • Virology
    • Genetics

    Background:

    • The 244-base-pair polyomavirus enhancer region is essential for viral DNA replication.
    • Previous research indicated that enhancer function is required for polyomavirus DNA replication.
    • The relationship between replication activation sequences and transcriptional enhancement sequences was not fully understood.

    Purpose of the Study:

    • To investigate the relationship between polyomavirus enhancer sequences involved in replication and transcription.
    • To functionally dissect the polyomavirus enhancer region and identify elements critical for replication activation.
    • To assess the dual role of identified sequence elements in both viral DNA replication and transcriptional enhancement.

    Main Methods:

    • Analysis of deletion mutants within the polyomavirus enhancer region.

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  • Synthesis of a specific DNA element (Element A, 26-mer) and its introduction into replication-deficient plasmids.
  • Assay of beta-globin mRNA levels to measure transcriptional enhancement activity of the synthesized element.

    • Main Results:

    • Four distinct sequence elements within the enhancer region were identified as involved in replication activation.
    • These identified elements exhibit functional redundancy and little sequence homology.
    • Multiple copies of a synthesized 26-mer (Element A) restored polyomavirus DNA replication and demonstrated transcriptional enhancer activity, correlating with copy number.

    Conclusions:

    • Specific sequence elements within the polyomavirus enhancer are critical for viral DNA replication.
    • These elements possess a dual function, acting as both replication activators and transcriptional enhancers.
    • The identified elements are functionally redundant, with multiple copies enhancing both processes.