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Related Experiment Videos

Accessory proteins impose site selectivity during ColE1 dimer resolution

A Guhathakurta1, I Viney, D Summers

  • 1Harvard University, Department of Biochemistry and Molecular Biology, Cambridge, Massachusetts 02138, USA.

Molecular Microbiology
|May 1, 1996
PubMed
Summary
This summary is machine-generated.

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Accessory proteins ArgR and PepA control plasmid DNA recombination, ensuring sites are monomeric. Their absence leads to multimerization, revealing their role in site selectivity and constraint.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • The cer-Xer system on plasmid ColE1 resolves dimers selectively via XerCD recombinase and accessory proteins ArgR and PepA.
  • The Escherichia coli dif site uses the same recombinase but lacks ArgR/PepA dependence and site selectivity, suggesting these proteins impose constraint.

Purpose of the Study:

  • To characterize novel "conditionally constrained" multimer resolution sites.
  • To investigate the role of ArgR and PepA in imposing site-specific constraint on DNA recombination.

Main Methods:

  • Characterization of conditionally constrained multimer resolution sites.
  • Analysis of plasmid multimerization in the presence and absence of ArgR and PepA.
  • Testing a mutant ArgR derivative (ArgR110) defective in cer-mediated dimer resolution.

Related Experiment Videos

  • Altering the distance between ArgR and XerCD binding sites.

    • Main Results:

    • Conditionally constrained sites maintain plasmid monomeric form with ArgR/PepA but lead to multimerization without them.
    • A mutant ArgR (ArgR110) still prevented multimerization, indicating a trans-acting blocking mechanism.
    • Altering the helical turn distance between binding sites abolished constraint, which was restored by a full helical turn.

    Conclusions:

    • ArgR and PepA are crucial for imposing constraint on multimer resolution sites, likely by blocking recombination in trans.
    • The precise helical phasing between accessory protein and recombinase binding sites is essential for site constraint.