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PMID: 6255431 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Inhibition of Eco RI action by polynucleotides. A characterization of the non-specific binding of the enzyme to DNA.

Nucleic acids research ·Vol. 8 ·No. 20 ·1980-10-24 ·Pages 4727-36

Langowski J, Pingoud A, Goppelt M, Maass G

Abstract

The cleavage of the plasmid pBR322 by the restriction endonuclease Eco RI has been studied in the presence of various polynucleotides and the double stranded octanucleotide d-(GGAATTCC) in order to clarify whether there is a preferential interaction of Eco RI with DNA sequences other than -GAATTC-. The steady state kinetic analysis shows that all polynucleotides investigated with the possible exception of poly-dG.poly-dC inhibit the cleavage competitively with Ki values in the range of 10(-4) to 10(-5) [M nucleotides]. The Ki of d-(GGAATTCC) is 1.5.10(-6) [M nucleotides], indicating that the specific binding is approx. 2 orders of magnitude stronger than non-specific binding.

MeSH Terms

DNA Restriction Enzymes/metabolism DNA, Bacterial Escherichia coli Kinetics Oligodeoxyribonucleotides/pharmacology Plasmids Polydeoxyribonucleotides/pharmacology Protein Binding Structure-Activity Relationship

Chemicals

DNA, Bacterial Oligodeoxyribonucleotides Polydeoxyribonucleotides DNA Restriction Enzymes

Authors & Affiliations

4 authors, click to expand affiliations / ORCID

Langowski J

Pingoud A

Goppelt M

Maass G

References (14)

14 references, click to expand

A deoxyribonucleic acid polymerase from Micrococcus luteus (Micrococcus lysodeikticus) isolated on deoxyribonucleic acid-cellulose.
J Biol Chem. 1968 Dec 10;243(23):6222-33 PMID: 5723464
Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form.
Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159-66 PMID: 4894690
Affinity chromatography of phosphofructokinase using Cibacron blue F3G-A.
J Chromatogr. 1972 Jun 28;69(1):209-14 PMID: 4261079
Diffusion controlled reaction rates in spheroidal geometry. Application to repressor--operator association and membrane bound enzymes.
Biophys Chem. 1974 Oct;2(3):255-63 PMID: 4474030
Non-specific DNA binding of genome regulating proteins as a biological control mechanism: I. The lac operon: equilibrium aspects.
Proc Natl Acad Sci U S A. 1974 Dec;71(12):4808-12 PMID: 4612528
Recognition mechanisms of DNA-specific enzymes.
Annu Rev Biochem. 1976;45:889-920 PMID: 134668
EcoRI endonuclease. Physical and catalytic properties of the homogenous enzyme.
J Biol Chem. 1976 Oct 10;251(19):5866-74 PMID: 786985
Superstructure of linear duplex DNA.
Proc Natl Acad Sci U S A. 1976 Nov;73(11):4125-9 PMID: 1069302
Kinetic studies on the cleavage of adenovirus DNA by restriction endonuclease Eco RI.
Nucleic Acids Res. 1976 Dec;3(12):3255-69 PMID: 794836
The effects of deoxyribonucleic acid secondary structure on tertiary structure.
Biochem J. 1976 Dec 1;159(3):615-20 PMID: 1008821
Restriction endonucleases.
CRC Crit Rev Biochem. 1976 Nov;4(2):123-64 PMID: 795607
Poly(dG).poly(dC) at neutral and alkaline pH: the formation of triple stranded poly(dG).poly(dG).poly(dC).
Nucleic Acids Res. 1978 Mar;5(3):1017-28 PMID: 25418
A hypothesis on a specific sequence-dependent conformation of DNA and its relation to the binding of the lac-repressor protein.
J Mol Biol. 1979 Jul 15;131(4):669-80 PMID: 513130
The interaction of the EcoRI restriction endonuclease with its substrate. A physico-chemical study employing natural and synthetic oligonucleotides and polynucleotides.
Eur J Biochem. 1980 Feb;104(1):101-7 PMID: 6245864

Article Info

Journal

Nucleic acids research

Abbr.

Nucleic Acids Res

ISSN

0305-1048

DOI

10.1093/nar/8.20.4727

Published

1980-10-24

Pages

4727-36

Language

English

Region

England

NLM ID

0411011

PMCID

PMC324382

Subset

External Links

PubMed source DOI: 10.1093/nar/8.20.4727 PMC Full Text View journal details

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Inhibition of Eco RI action by polynucleotides. A characterization of the non-specific binding of the enzyme to DNA.

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