Initiation of adenovirus DNA replication: Detection of covalent complexes between nucleotide and the 80-kilodalton terminal protein

M. D. Challberg, J. M. Ostrove, T. J. Kelly

Research output: Contribution to journalArticle

Abstract

We have previously shown that the 5'-terminal deoxycytidine residue of each nascent adenovirus 5 DNA strand synthesized in vitro is covalently linked to the 80-kilodalton (kd) terminal protein precursor via a phosphodiester bond to a serine residue in the protein. When extracts prepared from adenovirus 5-infected cells are incubated with [α-32P]dCTP as the only added deoxynucleoside triphosphate, complexes consisting of nucleotide covalently linked to the 80-kd protein can be detected. The nucleotide moieties present in such complexes include d(pC) and d(pCpA), the 5'-terminal nucleotide and dinucleotide of adenovirus 5 DNA, respectively, as well as some longer oligonucleotides. The formation of these complexes requires the presence of adenovirus DNA containing the attached 55-kd terminal protein and ATP. Extracts from H5ts125-infected cells which are defective in DNA replication catalyze complex formation to the same extent as extracts prepared from wild-type infected cells; thus, the presence of the adenovirus-coded 72-kd DNA-binding protein is apparently not required. Most, if not all, of the 80-kd protein-nucleotide complexes that are formed are noncovalently bound to the input viral DNA. These observations are consistent with the protein-priming model for the initiation of adenovirus DNA replication.

Original languageEnglish (US)
Pages (from-to)265-270
Number of pages6
JournalJournal of virology
Volume41
Issue number1
DOIs
StatePublished - 1982

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

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