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Easily unwound DNA sequences and hairpin structures in the Epstein-Barr virus origin of plasmid replication.
J Virol. 1993 May; 67(5):2707-15.JV

Abstract

The Epstein-Barr virus (EBV) origin of plasmid replication (oriP) includes two known cis-acting components, the dyad symmetry region and the family of repeats. We used P1 nuclease, a single-strand-specific endonuclease, to probe EBV oriP for DNA sequences that are intrinsically easy to unwind on a negatively supercoiled plasmid. Selective nuclease hypersensitivity was detected in the family of repeats on an oriP-containing plasmid and in the dyad symmetry region on a plasmid that lacks the family of repeats, indicating that the DNA in both cis-acting components is intrinsically easy to unwind. The hierarchy of nuclease hypersensitivity indicates that the family of repeats is more easily unwound than the dyad symmetry region, consistent with the hierarchy of helical stability predicted by computer analysis of the DNA sequence. A specific subset of the family of repeats is nuclease hypersensitive, and the DNA structure deduced from nucleotide-level analysis of the P1 nuclease nicks is a cruciform near a single-stranded bubble. The dyad symmetry region unwinds to form a broad single-stranded bubble containing hairpins in the 65-bp dyad sequence. We propose that the intrinsic ease of unwinding the dyad symmetry region, the actual origin of DNA replication, is an important component in the mechanism of initiation.

Authors+Show Affiliations

Molecular and Cellular Biology Department, Roswell Park Cancer Institute, Buffalo, New York 14263.No affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

8386273

Citation

Williams, D L., and D Kowalski. "Easily Unwound DNA Sequences and Hairpin Structures in the Epstein-Barr Virus Origin of Plasmid Replication." Journal of Virology, vol. 67, no. 5, 1993, pp. 2707-15.
Williams DL, Kowalski D. Easily unwound DNA sequences and hairpin structures in the Epstein-Barr virus origin of plasmid replication. J Virol. 1993;67(5):2707-15.
Williams, D. L., & Kowalski, D. (1993). Easily unwound DNA sequences and hairpin structures in the Epstein-Barr virus origin of plasmid replication. Journal of Virology, 67(5), 2707-15.
Williams DL, Kowalski D. Easily Unwound DNA Sequences and Hairpin Structures in the Epstein-Barr Virus Origin of Plasmid Replication. J Virol. 1993;67(5):2707-15. PubMed PMID: 8386273.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Easily unwound DNA sequences and hairpin structures in the Epstein-Barr virus origin of plasmid replication. AU - Williams,D L, AU - Kowalski,D, PY - 1993/5/1/pubmed PY - 1993/5/1/medline PY - 1993/5/1/entrez SP - 2707 EP - 15 JF - Journal of virology JO - J Virol VL - 67 IS - 5 N2 - The Epstein-Barr virus (EBV) origin of plasmid replication (oriP) includes two known cis-acting components, the dyad symmetry region and the family of repeats. We used P1 nuclease, a single-strand-specific endonuclease, to probe EBV oriP for DNA sequences that are intrinsically easy to unwind on a negatively supercoiled plasmid. Selective nuclease hypersensitivity was detected in the family of repeats on an oriP-containing plasmid and in the dyad symmetry region on a plasmid that lacks the family of repeats, indicating that the DNA in both cis-acting components is intrinsically easy to unwind. The hierarchy of nuclease hypersensitivity indicates that the family of repeats is more easily unwound than the dyad symmetry region, consistent with the hierarchy of helical stability predicted by computer analysis of the DNA sequence. A specific subset of the family of repeats is nuclease hypersensitive, and the DNA structure deduced from nucleotide-level analysis of the P1 nuclease nicks is a cruciform near a single-stranded bubble. The dyad symmetry region unwinds to form a broad single-stranded bubble containing hairpins in the 65-bp dyad sequence. We propose that the intrinsic ease of unwinding the dyad symmetry region, the actual origin of DNA replication, is an important component in the mechanism of initiation. SN - 0022-538X UR - https://www.unboundmedicine.com/medline/citation/8386273/Easily_unwound_DNA_sequences_and_hairpin_structures_in_the_Epstein_Barr_virus_origin_of_plasmid_replication_ L2 - http://jvi.asm.org/cgi/pmidlookup?view=long&pmid=8386273 DB - PRIME DP - Unbound Medicine ER -