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PMID: 6091053 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Changes in site specificity of single-strand-specific endonucleases on supercoiled PM2 DNA with temperature and ionic environment.

Nucleic acids research ·Vol. 12 ·No. 18 ·1984-09-25 ·Pages 7071-86

Kowalski D

Abstract

Mung bean nuclease sites in supercoiled PM2 DNA at neutral pH were located by linearizing the singly-nicked circular DNA product with venom phosphodiesterase followed by restriction endonuclease mapping. The locations of the sites varied with small changes in temperature and in concentration of NaC1 or magnesium ion. Different environmental changes which affect duplex stability in the same direction showed similar effects on the number of sites and in some cases resulted in identical cleavage patterns. Venom phosphodiesterase and P1 nuclease showed cleavage patterns similar to mung bean nuclease under the same environmental conditions and showed similar variations in cleavage patterns when environmental conditions were changed. Relaxed, closed-circular DNA was slowly cleaved at numerous sites whose locations did not vary with environment. Changes in site specificity are likely the result of environmental effects on the conformation of supercoiled DNA as opposed to effects on the single-strand-specific endonucleases themselves.

MeSH Terms
Bacteriophages Base Sequence DNA Restriction Enzymes/metabolism DNA, Single-Stranded DNA, Superhelical DNA, Viral Endonucleases/metabolism Kinetics Molecular Weight Osmolar Concentration Pseudomonas Single-Strand Specific DNA and RNA Endonucleases Substrate Specificity Temperature
Chemicals
DNA, Single-Stranded DNA, Superhelical DNA, Viral Endonucleases DNA Restriction Enzymes Single-Strand Specific DNA and RNA Endonucleases
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Kowalski D
References (42)
42 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/12.18.7071
Published
1984-09-25
Pages
7071-86
Language
English
Region
England
NLM ID
0411011
PMCID
PMC320143
Subset
IM
Grants
NIGMS NIH HHS · GM24950 · United States
NIGMS NIH HHS · GM30614 · United States
External Links
PubMed source DOI: 10.1093/nar/12.18.7071 PMC Full Text View journal details
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