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A repressor heterodimer binds to a chimeric operator.

M Hollis1, D Valenzuela, D Pioli

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

Proceedings of the National Academy of Sciences of the United States of America
|August 1, 1988
PubMed
Summary
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Researchers created a hybrid repressor protein that binds to new DNA sequences. Combining this hybrid with the original repressor formed a heterodimer recognizing asymmetric DNA operators, altering DNA-binding specificity.

Area of Science:

  • Molecular Biology
  • Genetics
  • Protein Engineering

Background:

  • The 434 repressor protein recognizes specific DNA operator sequences.
  • The P22 repressor protein recognizes different DNA operator sequences.
  • DNA-binding specificity is determined by the interaction between repressor proteins and DNA.

Purpose of the Study:

  • To engineer a novel DNA-binding protein with altered specificity.
  • To investigate the DNA-binding properties of a hybrid repressor protein.
  • To explore the formation and specificity of repressor heterodimers.

Main Methods:

  • Protein engineering by replacing solvent-exposed residues in the 434 repressor's DNA recognition helix with those from the P22 repressor.
  • Construction and characterization of the hybrid repressor 434R[alpha 3(P22R)].

Related Experiment Videos

  • Formation of heterodimers between 434 and 434R[alpha 3(P22R)] repressor monomers.
  • Analysis of DNA-binding specificity of the heterodimer to a chimeric P22/434 operator lacking two-fold rotational symmetry.

    • Main Results:

    • The hybrid protein 434R[alpha 3(P22R)] specifically binds to P22 operators.
    • Heterodimerization of 434 and 434R[alpha 3(P22R)] repressor monomers generates a new DNA-binding specificity.
    • The resulting heterodimer specifically recognizes a chimeric P22/434 operator that lacks two-fold rotational symmetry.

    Conclusions:

    • Protein engineering of DNA recognition helices can alter repressor-DNA binding specificity.
    • Heterodimerization of related repressor proteins can create novel DNA-binding specificities.
    • Asymmetric DNA operator sequences can be recognized by engineered repressor heterodimers.