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Identification of EBNA1 amino acid sequences required for the interaction of the functional elements of the Epstein-Barr virus latent origin of DNA replication.
J Virol. 1993 Jun; 67(6):3418-26.JV

Abstract

Epstein-Barr nuclear antigen 1 (EBNA1) activates DNA replication from the Epstein-Barr virus latent origin, oriP. This activation involves the direct interaction of EBNA1 dimers with multiple sites within the two noncontiguous functional elements of the origin, the family of repeats (FR) element and the dyad symmetry (DS) element. The efficient interaction of EBNA1 dimers bound to these two elements in oriP results in the formation of DNA loops in which the FR and DS elements are bound together through EBNA1. In order to elucidate the mechanism by which EBNA1 induces oriP DNA looping, we have investigated the DNA sequences and EBNA1 amino acids required for EBNA1-mediated DNA looping. Using a series of truncation mutants of EBNA1 produced in baculovirus and purified to apparent homogeneity, we have demonstrated that the EBNA1 DNA binding and dimerization domain is not sufficient to mediate oriP DNA looping and that an additional region(s) located between amino acids 346 and 450 is required. Single EBNA1-binding sites, separated by 930 bp of plasmid DNA, were also shown to support EBNA1-mediated looping, indicating that the formation of large EBNA1 complexes, such as those observed on oriP FR and DS elements, is not a requirement for looping.

Authors+Show Affiliations

Molecular Virology and Immunology Program, McMaster University, Hamilton, Ontario, Canada.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

8388506

Citation

Goldsmith, K, et al. "Identification of EBNA1 Amino Acid Sequences Required for the Interaction of the Functional Elements of the Epstein-Barr Virus Latent Origin of DNA Replication." Journal of Virology, vol. 67, no. 6, 1993, pp. 3418-26.
Goldsmith K, Bendell L, Frappier L. Identification of EBNA1 amino acid sequences required for the interaction of the functional elements of the Epstein-Barr virus latent origin of DNA replication. J Virol. 1993;67(6):3418-26.
Goldsmith, K., Bendell, L., & Frappier, L. (1993). Identification of EBNA1 amino acid sequences required for the interaction of the functional elements of the Epstein-Barr virus latent origin of DNA replication. Journal of Virology, 67(6), 3418-26.
Goldsmith K, Bendell L, Frappier L. Identification of EBNA1 Amino Acid Sequences Required for the Interaction of the Functional Elements of the Epstein-Barr Virus Latent Origin of DNA Replication. J Virol. 1993;67(6):3418-26. PubMed PMID: 8388506.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Identification of EBNA1 amino acid sequences required for the interaction of the functional elements of the Epstein-Barr virus latent origin of DNA replication. AU - Goldsmith,K, AU - Bendell,L, AU - Frappier,L, PY - 1993/6/1/pubmed PY - 1993/6/1/medline PY - 1993/6/1/entrez SP - 3418 EP - 26 JF - Journal of virology JO - J Virol VL - 67 IS - 6 N2 - Epstein-Barr nuclear antigen 1 (EBNA1) activates DNA replication from the Epstein-Barr virus latent origin, oriP. This activation involves the direct interaction of EBNA1 dimers with multiple sites within the two noncontiguous functional elements of the origin, the family of repeats (FR) element and the dyad symmetry (DS) element. The efficient interaction of EBNA1 dimers bound to these two elements in oriP results in the formation of DNA loops in which the FR and DS elements are bound together through EBNA1. In order to elucidate the mechanism by which EBNA1 induces oriP DNA looping, we have investigated the DNA sequences and EBNA1 amino acids required for EBNA1-mediated DNA looping. Using a series of truncation mutants of EBNA1 produced in baculovirus and purified to apparent homogeneity, we have demonstrated that the EBNA1 DNA binding and dimerization domain is not sufficient to mediate oriP DNA looping and that an additional region(s) located between amino acids 346 and 450 is required. Single EBNA1-binding sites, separated by 930 bp of plasmid DNA, were also shown to support EBNA1-mediated looping, indicating that the formation of large EBNA1 complexes, such as those observed on oriP FR and DS elements, is not a requirement for looping. SN - 0022-538X UR - https://www.unboundmedicine.com/medline/citation/8388506/Identification_of_EBNA1_amino_acid_sequences_required_for_the_interaction_of_the_functional_elements_of_the_Epstein_Barr_virus_latent_origin_of_DNA_replication_ L2 - http://jvi.asm.org/cgi/pmidlookup?view=long&pmid=8388506 DB - PRIME DP - Unbound Medicine ER -