Role of conserved residues in the activity of adenovirus preterminal protein.
Preterminal protein (pTP) is a component of the preinitiation complex which forms at the adenovirus origin of DNA replication and acts as the protein primer during DNA synthesis. In order to determine the role of various regions of the molecule a series of 18 mutations was introduced into conserved motifs of pTP which were predicted to be surface exposed, and the mutants expressed in insect cells using a baculovirus expression system. Their ability to initiate DNA replication was assessed and the effect the mutations have on the individual interactions which contribute to the formation of the pre-initiation complex was determined. Classes of mutants could be identified which were unable to bind DNA or interact with the adenovirus DNA polymerase, but one class of mutants retained these activities and yet failed to initiate DNA replication. These mutants therefore identify regions of pTP required for different aspects of adenovirus DNA replication.
Botting
Catherine H
CH
Centre for Biomolecular Sciences, School of Biology, University of St Andrews, Biomolecular Sciences Building, North Haugh, St Andrews, Fife KY16 9ST, UK1.
Hay
Ronald T
RT
Centre for Biomolecular Sciences, School of Biology, University of St Andrews, Biomolecular Sciences Building, North Haugh, St Andrews, Fife KY16 9ST, UK1.
D000260:Adenoviruses, Human; D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002460:Cell Line; D004261:DNA Replication; D004277:DNA, Single-Stranded; D004279:DNA, Viral / Q000096:biosynthesis; D005822:Genetic Vectors; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010750:Phosphoproteins / Q000235:genetics / Q000502:physiology*; D011485:Protein Binding; D011498:Protein Precursors / Q000235:genetics / Q000502:physiology*; D014764:Viral Proteins; D014779:Virus Replication
D004277:DNA, Single-Stranded; D004279:DNA, Viral; D010750:Phosphoproteins; D011498:Protein Precursors; D014764:Viral Proteins; C085476:pTP protein, adenovirus