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Related Experiment Videos

Interaction of bacteriophage lambda repressor with nonoperator DNA containing single-strand gaps.

R Sussman, J Resnick, K Calame

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

    Lambda repressor (ind+) binds strongly to gapped DNA, unlike other DNA forms or lambda repressor (ind-). This suggests DNA gaps are key targets for repressor binding during lambda induction.

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    Area of Science:

    • Molecular Biology
    • Genetics
    • Biochemistry

    Background:

    • Lambda repressor is crucial for regulating the lambda phage life cycle.
    • Previous models proposed repressor binding to host DNA lesions during lambda induction.

    Purpose of the Study:

    • To investigate the DNA binding preferences of lambda repressor variants.
    • To reevaluate models of lambda induction based on repressor-DNA interactions.

    Main Methods:

    • Direct binding assays were used to measure repressor affinity.
    • Purified lambda repressor (ind+) and (ind-) were tested against various DNA substrates.
    • Competition assays were performed using gapped nonoperator DNA.

    Main Results:

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  • Lambda repressor (ind+) exhibited high affinity for single-strand gapped DNA.
  • Binding affinity was significantly lower for double-stranded, nicked, or denatured DNA.
  • Lambda repressor (ind-) showed substantially reduced affinity for gapped DNA compared to ind+.

    • Conclusions:

    • Single-strand gaps in DNA are preferential binding sites for lambda repressor (ind+).
    • These findings challenge previous assumptions about repressor binding during lambda induction.
    • The data supports a model where repressor interacts with DNA structural anomalies.