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Related Experiment Videos

Two thymidylate synthetases in Bacillus subtilis.

J Neuhard, A R Price, L Schack

    Proceedings of the National Academy of Sciences of the United States of America
    |March 1, 1978
    PubMed
    Summary

    Bacillus subtilis has two thymidylate synthetases (TSaseA and TSaseB) at lower temperatures. Temperature shifts reveal distinct properties, aiding mutant selection for thyA and thyB genes.

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    Area of Science:

    • Molecular biology
    • Enzymology
    • Microbial genetics

    Background:

    • Bacillus subtilis utilizes thymidylate synthetases for DNA synthesis.
    • Two distinct thymidylate synthetases, TSaseA and TSaseB, are present under specific growth conditions.
    • The expression and function of these enzymes are linked to the thyA and thyB genes.

    Purpose of the Study:

    • To investigate the properties of two distinct thymidylate synthetases in Bacillus subtilis.
    • To explore the differential expression of TSaseA and TSaseB based on temperature.
    • To establish a selection method for thyA and thyB mutants.

    Main Methods:

    • Bacterial growth at different temperatures (below 37°C and at 46°C).
    • Physical separation and purification of TSaseA and TSaseB.
    • Enzyme activity assays to determine kinetic and physicochemical properties.

    Main Results:

    • Bacillus subtilis grown below 37°C contains both TSaseA and TSaseB.
    • At 46°C, only TSaseA activity is detected.
    • TSaseA and TSaseB were successfully separated and exhibited similar general activity requirements.
    • Significant differences were observed in the kinetic and physicochemical properties of TSaseA and TSaseB.

    Conclusions:

    • Temperature-dependent expression of thymidylate synthetases in Bacillus subtilis allows for straightforward positive selection of thyA and thyB mutants.
    • TSaseA and TSaseB are distinct enzymes with unique kinetic and physicochemical characteristics, despite sharing some functional similarities.

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