Replication and Resolution of Cloned Poxvirus Telomeres In Vivo Generates Linear Minichromosomes with Intact Viral Hairpin Termini

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dc.contributor.authorDeLange, A. M.
dc.contributor.authorReddy, M.
dc.contributor.authorScraba, D.
dc.contributor.authorUpton, C.
dc.contributor.authorMFadden, G.
dc.date.accessioned2015-07-06T18:06:54Z
dc.date.available2015-07-06T18:06:54Z
dc.date.copyright1986en_US
dc.date.issued1986
dc.description.abstractThe covalently closed terminal hairpins of the linear duplex-DNA genomes of the orthopoxvirus vaccinia and the leporipoxvirus Shope fibroma virus (SFV) have been cloned as imperfect palindromes within circular plasmids in yeast cells and recombination-deficient Escherichia coli. The viral telomeres inserted within these recombinant plasmids are equivalent to the inverted-repeat structures detected as telomeric replicative intermediates during poxvirus replication in vivo. Although the telomeres of vaccinia and SFV show little sequence homology, the termini from both viral genomes exist as AT-rich terminal hairpins with extrahelical bases and alternate "flip-flop" configurations. Using an in vivo replication assay in which circular plasmid DNA was transfected into poxvirus-infected cells, we demonstrated the efficient replication and resolution of the cloned imperfect palindromes to bona fide hairpin termini. The resulting linear minichromosomes, which were readily purified from transfected cells, were shown by restriction enzyme mapping and by electron microscopy to have intact covalently closed hairpin termini at both ends. In addition, staggered unidirectional deletion derivatives of both the cloned vaccinia and SFV telomeric palindromes localized an approximately 200-base-pair DNA region in which the sequence organization was highly conserved and which was necessary for the resolution event. These data suggest a conserved mechanism of the resolution of poxvirus telomeres.en_US
dc.description.reviewstatusRevieweden_US
dc.description.scholarlevelFacultyen_US
dc.description.sponsorshipA.D. and C.U. are supported by postdoctoral fellowship awards from the Alberta Heritage Foundation for Medical Research (AHFMR). G.M. is an AHFMR scholar. This work was funded by the Medical Research Council of Canada.en_US
dc.identifier.citationDeLange, AM et al. Replication and Resolution of Cloned Poxvirus Telomeres In Vivo Generates Linear Minichromosomes with Intact Viral Hairpin Termini J. Virology 59(2) 249-259en_US
dc.identifier.urihttp://jvi.asm.org/content/59/2/249.abstract
dc.identifier.urihttp://hdl.handle.net/1828/6292
dc.language.isoenen_US
dc.publisherAmerican Society for Microbiologyen_US
dc.subject.departmentDepartment of Biochemistry and Microbiology
dc.titleReplication and Resolution of Cloned Poxvirus Telomeres In Vivo Generates Linear Minichromosomes with Intact Viral Hairpin Terminien_US
dc.typeArticleen_US

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