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Related Experiment Videos

Cell surface-located deoxyribonucleic acid receptors in transformable pneumococci.

H Seto, R Lopez, A Tomasz

    Journal of Bacteriology
    |June 1, 1975
    PubMed
    Summary
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    Deoxyribonucleic acid (DNA) binding in competent pneumococci primarily occurs at the protoplast membrane. Unmasking sequestered DNA-binding sites in both competent and incompetent cells is key to transformation.

    Area of Science:

    • Microbiology
    • Molecular Biology
    • Genetics

    Background:

    • Bacterial competence facilitates genetic transformation.
    • Deoxyribonucleic acid (DNA) uptake is a crucial step in bacterial transformation.
    • Understanding DNA binding mechanisms in transformable bacteria like pneumococci is essential.

    Purpose of the Study:

    • To investigate the location and nature of deoxyribonucleic acid (DNA) binding sites in transformable pneumococci.
    • To elucidate the role of cell wall and membrane components in DNA binding.
    • To explore the regulation of DNA binding in both competent and incompetent bacterial states.

    Main Methods:

    • Adsorption of DNA to competent and incompetent pneumococci under various conditions (e.g., presence/absence of glucose).

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  • Treatment of bound DNA with shear forces and autolysin to assess binding stability.
  • Analysis of DNA binding in isolated cell wall-membrane complexes.
  • Induction of DNA binding in incompetent cells via mechanical or enzymatic damage.

    • Main Results:

    • At least 50% of adsorbed DNA binds to the protoplast membrane in competent cells.
    • Glucose presence stabilizes DNA binding to competent cells against shear and autolysin.
    • DNA binding sites are present in cell wall-membrane complexes but are lost during cell fractionation.
    • Damage or altered ionic conditions can unmask DNA-binding sites in incompetent cells, but without transformation.

    Conclusions:

    • The deoxyribonucleic acid (DNA)-binding site is likely a membrane component, possibly protein, stabilized by the cell wall.
    • Both competent and incompetent pneumococci possess sequestered DNA-binding sites that can be exposed.
    • The competence activator protein may induce transformation by unmasking these endogenous binding sites.