Interaction of bacteriophage T4 and T7 single-stranded DNA-binding proteins with DNA

Phys Biol. 2009 Jul 1;6(2):025002. doi: 10.1088/1478-3975/6/2/025002.

Abstract

Bacteriophages T4 and T7 are well-studied model replication systems, which have allowed researchers to determine the roles of many proteins central to DNA replication, recombination and repair. Here we summarize and discuss the results from two recently developed single-molecule methods to determine the salt-dependent DNA-binding kinetics and thermodynamics of the single-stranded DNA (ssDNA)-binding proteins (SSBs) from these systems. We use these methods to characterize both the equilibrium double-stranded DNA (dsDNA) and ssDNA binding of the SSBs T4 gene 32 protein (gp32) and T7 gene 2.5 protein (gp2.5). Despite the overall two-orders-of-magnitude weaker binding of gp2.5 to both forms of DNA, we find that both proteins exhibit four-orders-of-magnitude preferential binding to ssDNA relative to dsDNA. This strong preferential ssDNA binding as well as the weak dsDNA binding is essential for the ability of both proteins to search dsDNA in one dimension to find available ssDNA-binding sites at the replication fork.

Publication types

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

MeSH terms

  • Bacteriophage T4 / genetics
  • Bacteriophage T4 / metabolism*
  • Bacteriophage T7 / genetics
  • Bacteriophage T7 / metabolism*
  • DNA, Single-Stranded / chemistry
  • DNA, Single-Stranded / metabolism
  • DNA, Viral / chemistry
  • DNA, Viral / metabolism*
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / metabolism*
  • Nucleic Acid Denaturation
  • Protein Binding
  • Salts / chemistry

Substances

  • DNA, Single-Stranded
  • DNA, Viral
  • DNA-Binding Proteins
  • Salts