Replication of single-stranded DNA templates by primase-polymerase complexes of the yeast, Saccharomyces cerevisiae.
A partially purified primase-polymerase complex from the yeast, Saccharomyces cerevisiae, was capable of replicating a single stranded circular phage DNA into a replicative form with high efficiency. The primase-polymerase complex exhibited primase activity and polymerase activity on singly primed circular ssDNA as well as on gapped DNA. In addition, it was able to replicate an unprimed, single-stranded, circular phage DNA through a coupled primase-polymerase action. On Biogel A-O.5m filtration the primase-polymerase activities appeared in the void volume, demonstrating a mass of greater than 500 kilodaltons. Primase and various primase-polymerase complexes synthesized unique primers on single stranded DNA templates and the size distribution of primers was dependent on the structure of the DNA and the nature of the primase-polymerase assembly.
D000255:Adenosine Triphosphate / Q000502:physiology; D019915:DNA Primase; D004261:DNA Replication; D004277:DNA, Single-Stranded / Q000235:genetics*; D004259:DNA-Directed DNA Polymerase / Q000502:physiology*; D066298:In Vitro Techniques; D007700:Kinetics; D046911:Macromolecular Substances; D012316:RNA Nucleotidyltransferases / Q000502:physiology*; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D013698:Templates, Genetic
D004277:DNA, Single-Stranded; D046911:Macromolecular Substances; D000255:Adenosine Triphosphate; D019915:DNA Primase; D012316:RNA Nucleotidyltransferases; D004259:DNA-Directed DNA Polymerase