Domain organisation of the adenovirus preterminal protein.

A Webster, I Leith, Ronald Thomas Hay

Research output: Contribution to journalArticlepeer-review

19 Citations (Scopus)

Abstract

In adenovirus-infected cells, the virus-encoded preterminal protein and DNA polymerase form a heterodimer that is directly involved in initiation of DNA replication. Monoclonal, antibodies were raised against preterminal protein, and epitopes recognized by the antibodies were identified by using synthetic peptides. Partial proteolysis of preterminal protein reveals that it has a tripartite structure,,vith the three domains being separated by two protease-sensitive areas, located at sites processed by adenovirus protease. These areas of protease sensitivity are probably surface-exposed loops, as they are the sites, along with the C-terminal region of preterminal protein, recognized by the monoclonal antibodies. Preterminal protein is protected from proteolytic cleavage when bound to adenovirus DNA polymerase, suggesting either multiple contact points between the proteins or a DNA polymerase-induced conformational change in preterminal protein. Two of the preterminal protein-specific antibodies induced dissociation of the preterminal protein-adenovirus DNA polymerase heterodimer and inhibited initiation of adenovirus DNA replication in vitro. Antibodies binding close to the primary processing sites of adenovirus protease inhibited DNA binding, consistent with UV cross-linking results which reveal that an N-terminal, protease resistant domain of preterminal protein contacts DNA. Monoclonal antibodies recognizing epitopes within the C-terminal 60 amino acids of preterminal protein stimulate DNA binding, an effect mediated through a decrease in the dissociation rate constant. These results suggest that preterminal protein contains a large, noncontiguous surface required for interaction with DNA polymerase, an N-terminal DNA binding domain, and a C-terminal regulatory domain.

Original languageEnglish
Pages (from-to)539-547
Number of pages9
JournalJournal of Virology
Volume71
Publication statusPublished - Jan 1997

Keywords

  • DNA-BINDING PROTEIN
  • NUCLEAR FACTOR-I
  • HELIX-DESTABILIZING PROPERTIES
  • NF-KAPPA-B
  • TERMINAL PROTEIN
  • PREINITIATION COMPLEX
  • MONOCLONAL-ANTIBODIES
  • POLYMERASE COMPLEX
  • BACK MECHANISM
  • REPLICATION

Fingerprint

Dive into the research topics of 'Domain organisation of the adenovirus preterminal protein.'. Together they form a unique fingerprint.

Cite this