Domain structure and quaternary organization of the bacteriophage P22 Erf protein

J Mol Biol. 1983 Dec 25;171(4):401-18. doi: 10.1016/0022-2836(83)90037-2.

Abstract

The structure and activities of the recombination-promoting P22 Erf protein were examined in vitro. Treatment of the protein with elastase produces a stable amino-terminal fragment, consisting of amino acid residues 1 to (approximately) 136. We have purified this fragment, designated fragment B, to apparent homogeneity by gel filtration chromatography. Fragment B retains the oligomeric structure and single-stranded DNA binding specificity of intact Erf. It differs, however, in lacking the ability of intact Erf to bind single-stranded DNA into large aggregates following mild heat treatment of the protein. In addition, its binding to DNA may be weaker than that of intact Erf. Intact Erf sediments through a sucrose gradient as a discrete species with an apparent S20,w of approximately 11 X 7 S. Its sedimentation behavior is affected little, if at all, by concentration. Fragment B also sediments as a discrete species at approximately 10 X 4 S. In the electron microscope, intact Erf appears as rings, with 10 to 14 small projecting structures resembling the teeth of a gear. Fragment B is similar, except that it appears to lack the peripheral structures. From these observations, we conclude that Erf consists of at least two structurally and functionally distinct domains, and that it has a discrete ring-like oligomeric structure.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Amino Acids / analysis
  • Centrifugation, Density Gradient
  • Chromatography, Gel
  • DNA, Single-Stranded
  • Electrophoresis, Polyacrylamide Gel
  • Macromolecular Substances
  • Microscopy, Electron
  • Peptide Fragments / isolation & purification
  • Protein Conformation
  • Recombination, Genetic
  • Salmonella Phages / analysis*
  • Viral Proteins* / isolation & purification

Substances

  • Amino Acids
  • DNA, Single-Stranded
  • Macromolecular Substances
  • Peptide Fragments
  • Viral Proteins