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Deletion mapping the yeast TRP5 control region.

W S Moye, H Zalkin

    The Journal of Biological Chemistry
    |April 25, 1985
    PubMed
    Summary
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    The yeast TRP5 gene

    Area of Science:

    • Molecular Biology
    • Yeast Genetics
    • Gene Regulation

    Background:

    • The yeast TRP5 gene is part of the amino acid biosynthesis pathway.
    • Understanding gene regulation is crucial for metabolic engineering and synthetic biology.

    Purpose of the Study:

    • To investigate the promoter function and regulatory elements of the yeast TRP5 gene.
    • To identify specific DNA sequences responsible for the general control regulation of TRP5.

    Main Methods:

    • Construction of a TRP5-lacZ translational fusion in yeast.
    • Deletion analysis of the 5'-flanking region of the TRP5 gene.
    • Assay of beta-galactosidase activity to determine promoter function and regulation.

    Main Results:

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    • The TRP5-lacZ fusion demonstrated 4-5 fold derepression under lysine limitation, confirming regulation by general control.
    • Deletion mapping localized the TRP5 promoter to within 188 base pairs of the transcription start site.
    • Two separable promoter subregions, approximately -188 to -114 and -91 to -29, were identified, with the TATA box in the downstream region. Regulatory elements essential for general control overlap these regions, each containing a TGACT repeat.

    Conclusions:

    • The TRP5 gene promoter contains distinct regulatory elements essential for general control.
    • The identified promoter regions and TGACT repeats provide insights into the molecular mechanisms of amino acid biosynthesis regulation in yeast.