Mechanism of RNA primer removal by the RNase H activity of avian myeloblastosis virus reverse transcriptase

J. J. Champoux, Eli Gilboa, D. Baltimore

Research output: Contribution to journalArticle

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Abstract

The single-stranded DNA containing the Moloney murine leukemia virus origin for plus-strand synthesis was cloned in M13mp2 and used as a template for avian myeloblastosis virus reverse transcriptase in the presence of Moloney RNA which had been treated with pancreatic RNase A. The RNA pieces containing the polypyrine stretch near the plus-strand origin were processed, presumably by RNase H, to generate primers for DNA synthesis which initiated both at the correct origin site and at one nucleotide downstream from the correct site. Approximately 50% of the labeled DNA fragments synthesized under these conditions retained the priming RNA on their 5' ends. When the isolated fragments were hybridized back to the template DNA and again treated with the reverse transcriptase, all of the RNA was removed from the labeled DNA. By using 5'-end-labeled pancreatic RNase A-resistant fragments, it was possible to show that the RNA primers were removed intact. It appears from these results that the RNase H activity associated with the enzyme shows a preference for cutting at the junction between the RNA and DNA moieties of such complexes and therefore is ideally suited for removing RNA primers.

Original languageEnglish
Pages (from-to)686-691
Number of pages6
JournalJournal of Virology
Volume49
Issue number3
StatePublished - Jan 1 1984
Externally publishedYes

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Avian myeloblastosis virus
Avian Myeloblastosis Virus
Ribonuclease H
RNA-directed DNA polymerase
RNA-Directed DNA Polymerase
Pancreatic Ribonuclease
RNA
DNA
Moloney murine leukemia virus
DNA Primers
Single-Stranded DNA
Murine leukemia virus
single-stranded DNA
synthesis
DNA primers
Nucleotides
RNA primers
nucleotides
Enzymes

ASJC Scopus subject areas

  • Immunology

Cite this

Mechanism of RNA primer removal by the RNase H activity of avian myeloblastosis virus reverse transcriptase. / Champoux, J. J.; Gilboa, Eli; Baltimore, D.

In: Journal of Virology, Vol. 49, No. 3, 01.01.1984, p. 686-691.

Research output: Contribution to journalArticle

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