Interaction of EcoRII restriction and modification enzymes with synthetic DNA fragments. VI. The binding and cleavage of substrates containing nucleotide analogs.

Nucleic acids research ·Vol. 13 ·No. 24 ·1985-12-20 ·Pages 8983-98

Yolov AA, Vinogradova MN, Gromova ES, Rosenthal A, Cech D, Veiko VP, Metelev VG, Kosykh VG, Buryanov YI, Bayev AA

Abstract

The present study deals with the binding and cleavage by EcoRII endonuclease of concatemer DNA duplexes containing EcoRII recognition sites (formula; see text) in which dT is replaced by dU or 5-bromodeoxyuridine, or 5'-terminal dC in the dT-containing strand is methylated at position 5. The enzyme molecule is found to interact with the methyl group of the dT residue of the DNA recognition site and to be at least in proximity to the H5 atom of the 5'-terminal dC residue in dT-containing strand of this site. Modification of any of these positions exerts an equal effects on the cleavage of both DNA strands. Endonuclease EcoRII was found to bind the substrate specifically. At the same time modification of the bases in recognized sequence may result in the formation of unproductive, though stable, enzyme-substrate complexes.

MeSH Terms

Base Sequence DNA/metabolism DNA Restriction Enzymes/metabolism DNA-Binding Proteins/metabolism Deoxyribonucleases, Type II Site-Specific Kinetics Oligodeoxyribonucleotides/chemical synthesis,metabolism Protein Binding Structure-Activity Relationship

Chemicals

DNA-Binding Proteins Oligodeoxyribonucleotides DNA DNA Restriction Enzymes CCWGG-specific type II deoxyribonucleases Deoxyribonucleases, Type II Site-Specific

Authors & Affiliations

10 authors, click to expand affiliations / ORCID

Yolov A A

Vinogradova M N

Gromova E S

Rosenthal A

Cech D

Veiko V P

Metelev V G

Kosykh V G

Buryanov Y I

Bayev A A

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21 references, click to expand

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Article Info

Journal

Nucleic acids research

Abbr.

Nucleic Acids Res

ISSN

0305-1048

DOI

10.1093/nar/13.24.8983

Published

1985-12-20

Pages

8983-98

Language

English

Region

England

NLM ID

0411011

PMCID

PMC318966

Subset

External Links

PubMed source DOI: 10.1093/nar/13.24.8983 PMC Full Text View journal details

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Interaction of EcoRII restriction and modification enzymes with synthetic DNA fragments. VI. The binding and cleavage of substrates containing nucleotide analogs.

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