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Structural and functional alterations in microtubule protein from Chinese hamster ovary cell mutants

R A Keates, F Sarangi, V Ling

    Proceedings of the National Academy of Sciences of the United States of America
    |September 1, 1981
    PubMed

    Insights

    Chinese hamster ovary cells developed resistance to the antimicrotubule drug Colcemid. This resistance is linked to altered microtubule and tubulin properties, offering insights into microtubule assembly regulation.

    Area of Science:

    • Cell Biology
    • Biochemistry
    • Molecular Biology

    Background:

    • Antimicrotubule drugs like Colcemid are crucial for cancer therapy and cell biology research.
    • Understanding drug resistance mechanisms is vital for improving therapeutic efficacy and cell function studies.

    Purpose of the Study:

    • To investigate the molecular basis of Colcemid resistance in Chinese hamster ovary (CHO) cells.
    • To characterize the alterations in microtubule and tubulin properties associated with increasing drug resistance.

    Main Methods:

    • Selection of CHO cell mutants with increasing resistance to Colcemid.
    • Biochemical analysis of purified microtubule protein and tubulin.
    • In vitro assays to assess microtubule assembly dynamics and drug binding affinity.

    Main Results:

    • Mutant cells exhibited decreased Colcemid-binding affinity to tubulin.
    • Electrophoretic analysis revealed alterations in tubulin subcomponent patterns.
    • In vitro polymerization assays showed increased resistance to Colcemid inhibition and, in one mutant, a decreased critical concentration for assembly.

    Conclusions:

    • Colcemid resistance in CHO cells is associated with specific, stepwise alterations in microtubule and tubulin properties.
    • These characterized mutants provide valuable tools for studying the regulation of microtubule assembly in vivo.
    • The findings contribute to a deeper understanding of drug resistance mechanisms and microtubule dynamics.

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