Strand-Specific Supercoiled DNA-Protein Relaxation Complexes: Comparison of the Complexes of Bacterial Plasmids ColE1 and ColE2

AUTOR(ES)

Blair, D. G.

RESUMO

Certain bacterial plasmids can be isolated as unique complexes of supercoiled circular DNA and protein. These complexes are distinguished by the conversion of the supercoiled DNA to the relaxed or open-circular DNA form upon treatment with ionic detergents, proteases, or alkali. This report demonstrates that the open-circular DNA resulting from the pronase-induced relaxation of the complexes of colicinogenic factors E1 (ColE1) and E2 (ColE2) possesses a strand-specific break. In each case this break is found in the heavy strand of the DNA as defined by CsCl centrifugation in the presence of poly(U,G). In addition, the ColE1 and ColE2 complexes exhibit certain properties that are plasmid specific. Heat treatment, and to a lesser extent pronase treatment, inactivates the ColE2 complex, making it insensitive to agents that formerly were capable of inducing relaxation (conversion of the DNA to the open-circular form). In contrast, the ColE1 complex is not inactivated by these treatments. The potential role of these strand-specific relaxation complexes in DNA replication is discussed.

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