Does the specific recongnition of DNA by the restriction endonucleaseEcoRI involve a linear diffusion step? Investigation of the processivity of theEcoRI endonuclease
- 1 January 1983
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 11 (2) , 501-513
- https://doi.org/10.1093/nar/11.2.501
Abstract
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 8 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.Keywords
This publication has 15 references indexed in Scilit:
- Diffusion-driven mechanisms of protein translocation on nucleic acids. 1. Models and theoryBiochemistry, 1981
- Preparation of milligram amounts of 21 deoxyribonucleic acid restriction fragmentsBiochemistry, 1981
- Transient cleavage kinetics of the Eco RI restriction endonuclease measured in a pulsed quenchflow apparatus: enzyme concentration-dependent activity changeNucleic Acids Research, 1981
- The EcoRI restriction endonuclease with bacteriophage λ DNA. Kinetic studiesBiochemical Journal, 1980
- The Interaction of the EcoRI Restriction Endonuclease with its SubstrateEuropean Journal of Biochemistry, 1980
- Inhibition of Eco RI action by polynucleotides. A characterization of the non-specific binding of the enzyme to DNANucleic Acids Research, 1980
- Non-specific binding of restriction endonudease EcoR1 to DNANucleic Acids Research, 1980
- Gradient thin-layer chromatography of oligonucleotides on DEAE-cellulose: An alternative to homochromatographyAnalytical Biochemistry, 1979
- Rate enhancement by guided diffusion. Chain length dependence of repressor-operator association ratesBiophysical Chemistry, 1978
- Substrate dependence of the mechanism of EcoRI endonucleaseNucleic Acids Research, 1978