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Nucleic Acids Research, 1983, Vol. 11, No. 2 501-513
© 1983

ENZYMOLOGY

Does the specific recongnition of DNA by the restriction endonuclease EcoRI involve a linear diffusion step? Investigation of the processivity of the EcoRI endonuclease

J. Langowaski, J. Alves, A. Pingoud and G. Maass

Zentrum Biochemie, Abteilung Biophysikalishche Chemie, Medizinishche Hochschule Hannover Konstanty-Gutschow Str. 8, D-3000 Hannover, FRG

Received October 12, 1982. Revised November 25, 1982. Accepted December 22, 1982.

The time course of the EcoRI endonuclease catalysed cleavage of three substrates, two plasmid DNAs and one oligonucleotide, each with two EcoRI sites, was measured. The two plasmid DNAs with the EcoRI sites 318 and 96 base pairs apart are cut in a distributive fashion, while the oligonucleotide with the EcoRI sites 3 base pairs apart is cut in a partially processive manner.It is concluded that a linear diffusion of the EcoRI endonuclease on its substrate across long stretches of DNA is not likely to be operative during the recognition process. Microscopic dissociation-reassociation processes, however, increase the probability of the enzyme to attack further sites located in the immediate vicinity of a given site.


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