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PMID: 6302683 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Conformational flexibility of junctions between contiguous B- and Z-DNAs in supercoiled plasmids.

Singleton CK, Klysik J, Wells RD

Abstract

The junctions between contiguous right- and left-handed helical DNA segments in supercoiled plasmids are shown to exhibit both sequence- and superhelical-dependent conformational flexibility. The nonalternating (purine-pyrimidine) tetranucleotide d(G-A-T-C), which is imbedded within a (dC-dG) tract and exists as a junction at low-density values, may be adopting a left-handed helix under the influence of higher negative supercoiling. The effect of supercoiling on the transition from a right- to a left-handed state for (dC-dG)n inserts, where n = 16, 13, and 5, also was evaluated. Only moderate amounts of superhelicity (-0.03 to -0.075) are required for formation and stabilization of left-handed DNA even for the (dC-dG) region of 10 dC X dG base pairs. Finally, we demonstrate that junction formation within a BamHI restriction site causes a drastic inhibition (greater than 80%) of cleavage by this restriction endonuclease.

MeSH Terms
DNA Restriction Enzymes/metabolism DNA, Superhelical/analysis Deoxyribonuclease BamHI Deoxyribonuclease EcoRI Endonucleases/metabolism Nucleic Acid Conformation Plasmids Polydeoxyribonucleotides/analysis Single-Strand Specific DNA and RNA Endonucleases
Chemicals
DNA, Superhelical Polydeoxyribonucleotides poly(dC-dG) Endonucleases DNA Restriction Enzymes Deoxyribonuclease BamHI Deoxyribonuclease EcoRI Single-Strand Specific DNA and RNA Endonucleases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Singleton C K
Klysik J
Wells R D
References (17)
17 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
1983-05-00
Pages
2447-51
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC393842
Subset
IM
Grants
NCI NIH HHS · CA20279 · United States
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