In vitro transcription of reconstituted 35S RNA·tRNAtrp template·primer complexes by the avian oncornavirus DNA polymerase Effect of temperature on the size of the DNA transcripts

J. P. Leis*, R. E. Smith, P. Dierks, J. T. Parsons, M. S. Collett, A. J. Faras

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

The synthesis of long DNA transcripts from avian sarcoma virus 35S RNA·tRNAtrp template·primer complexes in reconstituted reactions in vitro is temperature dependent. At 15° DNA transcripts accumulate that are 100 nucleotides in length (DNA100), whereas at 30° DNA transcripts ranging to 3000 nucleotides in length (DNA>100) are synthesized from these template·primer complexes. By hybridization analysis we have demonstrated that DNA100 synthesized at 15° contains nucleotide sequences complementary to the 5′ end of the viral RNA genome whereas longer DNA transcripts synthesized at 30° contain nucleotide sequences complementary to the 3′ terminus of the viral genome. Pulse-chase experiments indicate that the 5′-located DNA100 transcripts are precursors of the larger DNA transcripts. These data are consistent with the elongation of 5′ terminally located DNA transcripts at the 3′ end of the viral genome.

Original languageEnglish (US)
Pages (from-to)28-42
Number of pages15
JournalVirology
Volume85
Issue number1
DOIs
StatePublished - Mar 1978

ASJC Scopus subject areas

  • Virology

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