title: Identification of a cis-element that determines autonomous DNA replication in eukaryotic cells.
abstract: A 36-bp human consensus sequence (CCTMDAWKSGBYTSMAAWTWBCMYTTRSCAAATTCC) is capable of supporting autonomous replication of a plasmid after transfection into eukaryotic cells. After transfection and in vitro DNA replication, replicated plasmid DNA containing a mixture of oligonucleotides of this consensus was found to reiterate the consensus. Initiation of DNA replication in vitro occurs within the consensus. One version, A3/4, in pYACneo, could be maintained under selection in HeLa cells, unrearranged and replicating continuously for >170 cell doublings. Stability of plasmid without selection was high (> or =0.9/cell/generation). Homologs of the consensus are found consistently at mammalian chromosomal sites of initiation and within CpG islands. Versions of the consensus function as origins of DNA replication in normal and malignant human cells, immortalized monkey and mouse cells, and normal cow, chicken, and fruit fly cells. Random mutagenesis studies suggest an internal 20-bp consensus sequence of the 36 bp may be sufficient to act as a core origin element. This cis-element consensus sequence is an opportunity for focused analyses of core origin elements and the regulation of initiation of DNA replication.
authors.affiliation: McGill Cancer Centre, McGill University, Montreal, Quebec H3G 1Y6.
authors.forename: Gerald B
authors.initials: GB
authors.lastname: Price
authors.forename: Minna
authors.initials: M
authors.lastname: Allarakhia
authors.forename: Nandini
authors.initials: N
authors.lastname: Cossons
authors.forename: Torsten
authors.initials: T
authors.lastname: Nielsen
authors.forename: Maria
authors.initials: M
authors.lastname: Diaz-Perez
authors.forename: Paula
authors.initials: P
authors.lastname: Friedlander
authors.forename: Liang
authors.initials: L
authors.lastname: Tao
authors.forename: Maria
authors.initials: M
authors.lastname: Zannis-Hadjopoulos
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D016384:Consensus Sequence; D018899:CpG Islands; D017931:DNA Primers; D004261:DNA Replication; D004330:Drosophila; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D016296:Mutagenesis
chemical_list: D017931:DNA Primers