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Short and long range activation by the SV40 enhancer.

B Wasylyk, C Wasylyk, P Chambon

    Nucleic Acids Research
    |July 25, 1984
    PubMed
    Summary

    Adding DNA between the SV40 enhancer and promoters significantly reduces transcription activation. This short-range effect diminishes with increasing DNA length, impacting both SV40 early and conalbumin promoters.

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    Area of Science:

    • Molecular Biology
    • Gene Regulation
    • Biochemistry

    Background:

    • Enhancers are crucial regulatory DNA elements that increase gene transcription.
    • The SV40 enhancer is a well-studied model system for investigating enhancer function.
    • Promoters are DNA sequences where transcription initiation occurs.

    Purpose of the Study:

    • To investigate the effect of interposing DNA sequences on enhancer-promoter communication.
    • To determine the distance-dependent relationship between the SV40 enhancer and different promoter elements.
    • To analyze the impact of DNA length and orientation on transcriptional activation.

    Main Methods:

    • Construction of recombinant DNA molecules with varying lengths of interposed DNA between the SV40 enhancer and promoter elements (SV40 early and conalbumin).
    • Assay of transcriptional activation using reporter gene expression.
    • Analysis of enhancer-promoter interactions in different orientations.

    Main Results:

    • Transcriptional activation by the SV40 enhancer decreased in a biphasic manner with increasing lengths of interposed DNA.
    • Short DNA inserts (up to 150 bp) reduced activation to less than 10% of maximum.
    • Longer inserts (275 bp, 650 bp, 3737 bp) further decreased activation, but transcription remained elevated compared to enhancerless controls.

    Conclusions:

    • The functional communication between the SV40 enhancer and promoters is sensitive to the intervening DNA sequence length.
    • A short-range effect significantly impacts enhancer activity, suggesting spatial constraints or sequence-specific interactions.
    • Despite distance-dependent attenuation, the SV40 enhancer maintains a significant positive effect on transcription initiation.

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