Localization of bacterial DNA polymerase: evidence for a factory model of replication

Science. 1998 Nov 20;282(5393):1516-9. doi: 10.1126/science.282.5393.1516.

Abstract

Two general models have been proposed for DNA replication. In one model, DNA polymerase moves along the DNA (like a train on a track); in the other model, the polymerase is stationary (like a factory), and DNA is pulled through. To distinguish between these models, we visualized DNA polymerase of the bacterium Bacillus subtilis in living cells by the creation of a fusion protein containing the catalytic subunit (PolC) and green fluorescent protein (GFP). PolC-GFP was localized at discrete intracellular positions, predominantly at or near midcell, rather than being distributed randomly. These results suggest that the polymerase is anchored in place and thus support the model in which the DNA template moves through the polymerase.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Bacillus subtilis / enzymology
  • Bacillus subtilis / growth & development
  • Bacillus subtilis / metabolism*
  • Chromosomes, Bacterial / metabolism*
  • DNA Replication*
  • DNA, Bacterial / biosynthesis*
  • DNA-Directed DNA Polymerase / analysis*
  • DNA-Directed DNA Polymerase / metabolism
  • Green Fluorescent Proteins
  • Luminescent Proteins
  • Models, Biological
  • Recombinant Fusion Proteins / metabolism
  • Replication Origin*
  • Templates, Genetic

Substances

  • DNA, Bacterial
  • Luminescent Proteins
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins
  • DNA-Directed DNA Polymerase