Effect of ribonucleotides embedded in a DNA template on HIV-1 reverse transcription kinetics and fidelity

Waaqo Daddacha, Erin Noble, Laura A. Nguyen, Edward M. Kennedy, Baek Kim

Research output: Contribution to journalArticle

Abstract

HIV-1 reverse transcriptase (RT) frequently incorporates ribonucleoside triphosphates (rNTPs) during proviral DNA synthesis, particularly under the limited dNTP conditions found in macrophages. We investigated the mechanistic impacts of an rNMP embedded in DNA templates on HIV-1 RT-mediated DNA synthesis. We observed that the template-embedded rNMP induced pausing of RT and delayed DNA synthesis kinetics at low macrophage dNTP concentrations but not at high T cell dNTP concentrations. Although the binding affinity of RT to the rNMP-containing template-primer was not altered, the dNTP incorporation kinetics of RT were significantly reduced at one nucleotide upstream and downstream of the rNMP site, leading to pause sites. Finally, HIV-1 RT becomes more error-prone at rNMP sites with an elevated mismatch extension capability but not enhanced misinsertion capability. Together these data suggest that rNMPs embedded in DNA templates may influence reverse transcription kinetics and impact viral mutagenesis in macrophages.

Original languageEnglish (US)
Pages (from-to)12522-12532
Number of pages11
JournalJournal of Biological Chemistry
Volume288
Issue number18
DOIs
StatePublished - May 3 2013
Externally publishedYes

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Ribonucleotides
Transcription
Reverse Transcription
HIV-1
Macrophages
RNA-Directed DNA Polymerase
Kinetics
DNA
Ribonucleosides
Mutagenesis
T-cells
Nucleotides
T-Lymphocytes
Human immunodeficiency virus 1 reverse transcriptase

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Effect of ribonucleotides embedded in a DNA template on HIV-1 reverse transcription kinetics and fidelity. / Daddacha, Waaqo; Noble, Erin; Nguyen, Laura A.; Kennedy, Edward M.; Kim, Baek.

In: Journal of Biological Chemistry, Vol. 288, No. 18, 03.05.2013, p. 12522-12532.

Research output: Contribution to journalArticle

Daddacha, Waaqo ; Noble, Erin ; Nguyen, Laura A. ; Kennedy, Edward M. ; Kim, Baek. / Effect of ribonucleotides embedded in a DNA template on HIV-1 reverse transcription kinetics and fidelity. In: Journal of Biological Chemistry. 2013 ; Vol. 288, No. 18. pp. 12522-12532.
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