title: Segregation of a single outboard left-end origin is essential for the viability of parvovirus minute virus of mice.
abstract: During DNA replication, the hairpin telomeres of Minute Virus of Mice (MVM) are extended and copied to create imperfectly palindromic duplex junction sequences that bridge adjacent genomes in concatameric replicative-form DNA. These are resolved by the viral initiator protein, NS1, but mechanisms employed at the two telomeres differ. Left-end:left-end junctions are resolved asymmetrically at a single site, OriLTC, by NS1 acting in concert with a host factor, parvovirus initiation factor (PIF). Replication segregates doublet and triplet sequences, initially present as unpaired nucleotides in the bubble region of the left-end hairpin stem, to either side of the junction. These act as spacers between the NS1 and PIF binding sites, and their asymmetric distribution sets up active (OriLTC) and inactive (OriLGAA) forms of OriL. We used a reverse genetic approach to disrupt this asymmetry and found that neither opposing doublets nor triplets in the hairpin bubble were tolerated. Viable mutants were isolated at low frequency and found to contain second-site mutations that either restored the asymmetry or crippled one PIF binding site. These mutations either inactivated the inboard or activated the outboard form of OriL, a polarity that strongly suggests that, in the genus Parvovirus, an active inboard OriL is lethal.
authors.affiliation: Department of Laboratory Medicine, Yale University Medical School, 333 Cedar St., CB408, New Haven, CT 067510, USA.
authors.forename: Erik
authors.initials: E
authors.lastname: Burnett
authors.forename: Susan F
authors.initials: SF
authors.lastname: Cotmore
authors.forename: Peter
authors.initials: P
authors.lastname: Tattersall
mesh_terms: D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D004261:DNA Replication / Q000235:genetics; D004279:DNA, Viral / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D005804:Genes, Lethal; D005814:Genes, Viral; D051379:Mice; D008915:Minute Virus of Mice / Q000235:genetics* / Q000502:physiology; D008969:Molecular Sequence Data; D009154:Mutation; D009690:Nucleic Acid Conformation; D010641:Phenotype; D018741:Replication Origin; D016615:Telomere / Q000235:genetics / Q000378:metabolism
chemical_list: D004279:DNA, Viral