RNA primer handoff in bacteriophage T4 DNA replication: the role of single-stranded DNA-binding protein and polymerase accessory proteins

J Biol Chem. 2008 Aug 15;283(33):22838-46. doi: 10.1074/jbc.M802762200. Epub 2008 May 28.


In T4 phage, coordinated leading and lagging strand DNA synthesis is carried out by an eight-protein complex termed the replisome. The control of lagging strand DNA synthesis depends on a highly dynamic replisome with several proteins entering and leaving during DNA replication. Here we examine the role of single-stranded binding protein (gp32) in the repetitive cycles of lagging strand synthesis. Removal of the protein-interacting domain of gp32 results in a reduction in the number of primers synthesized and in the efficiency of primer transfer to the polymerase. We find that the primase protein is moderately processive, and this processivity depends on the presence of full-length gp32 at the replication fork. Surprisingly, we find that an increase in the efficiency of primer transfer to the clamp protein correlates with a decrease in the dissociation rate of the primase from the replisome. These findings result in a revised model of lagging strand DNA synthesis where the primase remains as part of the replisome after each successful cycle of Okazaki fragment synthesis. A delay in primer transfer results in an increased probability of the primase dissociating from the replication fork. The interplay between gp32, primase, clamp, and clamp loader dictates the rate and efficiency of primer synthesis, polymerase recycling, and primer transfer to the polymerase.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteriophage T4 / enzymology
  • Bacteriophage T4 / genetics*
  • DNA Replication*
  • DNA, Single-Stranded / genetics*
  • DNA, Viral / genetics*
  • DNA-Binding Proteins / metabolism*
  • DNA-Directed DNA Polymerase / genetics*
  • DNA-Directed DNA Polymerase / metabolism
  • DNA-Directed RNA Polymerases / genetics*
  • DNA-Directed RNA Polymerases / metabolism
  • RNA / metabolism*
  • Viral Proteins / genetics
  • Viral Proteins / metabolism


  • DNA, Single-Stranded
  • DNA, Viral
  • DNA-Binding Proteins
  • RNA primers
  • Viral Proteins
  • RNA
  • DNA-Directed RNA Polymerases
  • DNA-Directed DNA Polymerase