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PMID: 6340109 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Perfect palindromic lac operator DNA sequence exists as a stable cruciform structure in supercoiled DNA in vitro but not in vivo.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 80 ·No. 7 ·1983-04-00 ·Pages 1797-801

Sinden RR, Broyles SS, Pettijohn DE

Abstract

A perfect palindromic 66-base pair (bp) DNA sequence derived from the lac operator and cloned into plasmid pMB9 [Betz, J. L. & Sadler, J. R. (1981) Gene 13, 1-12] can exist in a 66-bp linear form or as two 33-bp cruciform arms. The fraction of the sequence in the cruciform depends on the superhelical density of the plasmid DNA. Relaxed DNA contains no cruciforms. The palindrome in the cruciform structure is cut by EcoRI endonuclease at the base of the cruciform arms, releasing 33-bp fragments; when in the linear form only 66-bp fragments are produced. The cruciform structure is fixed by trimethylpsoralen crosslinks in the cruciform arms. This together with the EcoRI cutting provides an assay for the cruciform structures in the DNA of living cells. Using this assay we show that the cruciform structure rarely if ever exists in vivo, but after DNA isolation greater than 90% of the sequence is in cruciforms. Results suggest that the plasmid DNA as organized in vivo either lacks sufficient torsional tension to form this cruciform or the palindrome is restrained in the linear form by other bound molecules.

MeSH Terms
Base Sequence DNA, Bacterial DNA, Superhelical Escherichia coli/genetics Furocoumarins Lac Operon Nucleic Acid Conformation Operon Plasmids
Chemicals
DNA, Bacterial DNA, Superhelical Furocoumarins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Sinden R R
Broyles S S
Pettijohn D E
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34 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
DOI
10.1073/pnas.80.7.1797
Published
1983-04-00
Pages
1797-801
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC393696
Subset
IM
Grants
NIGMS NIH HHS · GM 07703-2 · United States
NIGMS NIH HHS · GM 18243-11 · United States
External Links
PubMed source DOI: 10.1073/pnas.80.7.1797 PMC Full Text View journal details
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