Targeting of DNA polymerase to the adenovirus origin of DNA replication by interaction with nuclear factor I

Proc Natl Acad Sci U S A. 1994 Nov 22;91(24):11537-41. doi: 10.1073/pnas.91.24.11537.


Efficient initiation by the DNA polymerase of adenovirus type 2 requires nuclear factor I (NFI), a cellular sequence-specific transcription factor. Three functions of NFI--dimerization, DNA binding, and activation of DNA replication--are colocalized within the N-terminal portion of the protein. To define more precisely the role of NFI in viral DNA replication, a series of site-directed mutations within the N-terminal domain have been generated, thus allowing the separation of all three functions contained within this region. Impairment of the dimerization function prevents sequence-specific DNA binding and in turn abolishes the NFI-mediated activation of DNA replication. NFI DNA-binding activity, although necessary, is not sufficient to activate the initiation of adenovirus replication. A distinct class of NFI mutations that abolish the recruitment of the viral DNA polymerase to the origin also prevent the activation of replication. Thus, a direct interaction of NFI with the viral DNA polymerase complex is required to form a stable and active preinitiation complex on the origin and is responsible for the activation of replication by NFI.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviruses, Human / genetics*
  • Amino Acid Sequence
  • Base Sequence
  • DNA Primers / chemistry
  • DNA Replication
  • DNA-Binding Proteins / metabolism*
  • DNA-Directed DNA Polymerase / metabolism*
  • In Vitro Techniques
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • NFI Transcription Factors
  • Protein Binding
  • Structure-Activity Relationship
  • Transcription Factors / metabolism*
  • Viral Proteins / metabolism*
  • Virus Replication*


  • DNA Primers
  • DNA-Binding Proteins
  • NFI Transcription Factors
  • Transcription Factors
  • Viral Proteins
  • terminal protein, adenovirus
  • transcription factor nuclear factor 1
  • DNA-Directed DNA Polymerase