Structure and function of the adenovirus origin of replication

Dan R. Rawlins, Philip J. Rosenfeld, Ronald J. Wides, Mark D. Challberg, Thomas J. Kelly

Research output: Contribution to journalArticlepeer-review

92 Scopus citations

Abstract

Efficient initiation of adenovirus DNA replication requires the presence of specific terminal nucleotide sequences that collectively constitute the viral origin of replication. Using plasmids with deletions or base substitutions in a cloned segment of DNA derived from the terminus of the adenovirus 2 genome, we have demonstrated that the origin contains two functionally distinct regions. The first 18 bp of the viral genome are sufficient to support a limited degree of initiation. However, the presence of a sequence in the region between nucleotides 19 and 67 greatly enhances the efficiency of the initiation reaction. This region contains a specific binding site for a protein present in uninfected cells (KD = 2 × 10-11 M). The bound protein protects the DNA segment between base pairs 19 and 43 from attack by DNAase I. Studies with deletion mutants indicate that binding of the cellular protein is responsible for the enhancement of initiation.

Original languageEnglish
Pages (from-to)309-319
Number of pages11
JournalCell
Volume37
Issue number1
DOIs
StatePublished - May 1984
Externally publishedYes

Bibliographical note

Funding Information:
We thank Rene Crombie and Patricia Morrow for technical assistance and Dr. Gary Ketner for helpful discussions. We are grateful to Dr. M. Green for providing antibody to the adenovirus terminal protein. We are also grateful to Marcia Bolanowski for excellent secreiarial assistance in preparing the manuscript. The work described in this paper was supported by grants from the Natlonal Institutes of Health.

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