Protein-protein interactions in the bacteriophage T4 replisome. The leading strand holoenzyme is physically linked to the lagging strand holoenzyme and the primosome

J Biol Chem. 2003 Jan 31;278(5):3145-52. doi: 10.1074/jbc.M209858200. Epub 2002 Nov 9.

Abstract

The bacteriophage T4 replication complex is composed of eight proteins that function together to replicate DNA. This replisome can be broken down into four basic units: a primosome composed of gp41, gp61, and gp59; a leading strand holoenzyme composed of gp43, gp44/62, and gp45; a lagging strand holoenzyme; and a single strand binding protein polymer. These units interact further to form the complete replisome. The leading and lagging strand polymerases are physically linked in the presence of DNA or an active replisome. The region of interaction was mapped to an extension of the finger domain, such that Cys-507 of one subunit is in close proximity to Cys-507 of a second subunit. The leading strand polymerase and the primosome also associate, such that gp59 mediates the contact between the two complexes. Binding of gp43 to the primosome complex causes displacement of gp32 from the gp59.gp61.gp41 primosome complex. The resultant species is a complex of proteins that may allow coordinated leading and lagging strand synthesis, helicase DNA unwinding activity, and polymerase nucleotide incorporation.

Publication types

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

MeSH terms

  • Bacteriophage T4 / genetics*
  • Bacteriophage T4 / physiology*
  • Base Sequence
  • Cysteine
  • DNA Replication
  • DNA, Viral / chemistry*
  • DNA, Viral / genetics
  • DNA, Viral / metabolism*
  • Energy Transfer
  • Models, Molecular
  • Molecular Sequence Data
  • Protein Conformation
  • Spectrometry, Fluorescence
  • Templates, Genetic
  • Viral Proteins / chemistry*
  • Viral Proteins / metabolism
  • Virus Replication / physiology

Substances

  • DNA, Viral
  • Viral Proteins
  • Cysteine