Relaxation complexes of plasmid DNA and protein. II. Characterization of the proteins associated with the unrelaxed and relaxed complexes of plasmid ColE1.
The proteins of the DNA-protein relaxation complex of plasmid ColE1 were labeled with [3H]leucine by growth of ColE1 containing Escherichia coli cells in the presence of this radioactive labeled amino acid. Three major [3H]leucine-labeled proteins are found associated with the supercoiled DNA in the ColE1 relaxation complex. The molecular weights of these proteins, determined by sodium dodecyl sulfate-acrylamide gel electrophoresis, are 60,000, 16,000, and 11,000, respectively. Induction of relaxation of the supercoiled DNA by treatment of the complex with sodium dodecyl sulfate results in a dissociation of the two smaller proteins from the DNA. The 60,000 protein, however, remains associated specifically with the nicked strand of the open circular DNA. The strand-specific association of this protein with the relaxed DNA resists heat denaturation of the DNA, sedimentation through an alkaline (pH 12.5) sucrose gradient, and centrifugation to equilibrium in an alkaline (pH 12.5) CsCl gradient.
D001426:Bacterial Proteins / Q000302:isolation & purification; D001665:Binding Sites; D002499:Centrifugation, Density Gradient; D004269:DNA, Bacterial / Q000302:isolation & purification; D003851:Deoxyribonucleases; D004926:Escherichia coli / Q000032:analysis*; D005111:Extrachromosomal Inheritance; D046911:Macromolecular Substances; D008970:Molecular Weight; D009690:Nucleic Acid Conformation; D010957:Plasmids; D011402:Pronase; D011485:Protein Binding; D011487:Protein Conformation; D012967:Sodium Dodecyl Sulfate
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D046911:Macromolecular Substances; D012967:Sodium Dodecyl Sulfate; D003851:Deoxyribonucleases; D011402:Pronase