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Retroviruses have a single-stranded RNA genome that undergoes a special form of replication. Once the retrovirus has entered the host cell, an enzyme called reverse transcriptase synthesizes double-stranded DNA from the retroviral RNA genome. This DNA copy of the genome is then integrated into the host’s genome inside the nucleus via an enzyme called integrase. Consequently, the retroviral genome is transcribed into RNA whenever the host’s genome is transcribed, allowing the...
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Updated: Mar 31, 2026

Determining 3'-Termini and Sequences of Nascent Single-Stranded Viral DNA Molecules during HIV-1 Reverse Transcription in Infected Cells
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Determining 3'-Termini and Sequences of Nascent Single-Stranded Viral DNA Molecules during HIV-1 Reverse Transcription in Infected Cells

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Mechanistic and Kinetic Differences between Reverse Transcriptases of Vpx Coding and Non-coding Lentiviruses.

Gina M Lenzi1, Robert A Domaoal1, Dong-Hyun Kim2

  • 1From the Center for Drug Discovery, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia 30322.

The Journal of Biological Chemistry
|October 21, 2015
PubMed
Summary

Human Immunodeficiency Virus Type 1 (HIV-1) reverse transcriptase (RT) has evolved a faster polymerization step to overcome low dNTP concentrations in macrophages, unlike HIV-2/SIV which uses Vpx protein to degrade SAMHD1. This difference explains HIV-1

Keywords:
DNA replicationSAMHD1VpxdNTPenzyme kineticslentivirusmacrophagereverse transcription

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Determining 3'-Termini and Sequences of Nascent Single-Stranded Viral DNA Molecules during HIV-1 Reverse Transcription in Infected Cells
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Area of Science:

  • Virology
  • Molecular Biology
  • Biochemistry

Background:

  • Lentiviruses like HIV-1, HIV-2, and SIV exhibit differential replication kinetics in non-dividing macrophages.
  • HIV-1 replication is delayed in macrophages, while HIV-2/SIV replicates rapidly due to the Vpx protein.
  • Vpx counteracts the host restriction factor SAMHD1, a dNTPase that depletes deoxynucleotide triphosphates (dNTPs) essential for reverse transcription.

Purpose of the Study:

  • To kinetically analyze and compare the reverse transcriptases (RTs) of Vpx-coding (HIV-2/SIV) and Vpx-non-coding (HIV-1) lentiviruses.
  • To investigate how differences in RT kinetics relate to viral replication strategies in low-dNTP environments like macrophages.

Main Methods:

  • Pre-steady-state kinetic analysis of RTs from HIV-1, HIV-2, and SIV.
  • Assessed polymerization rate (kpol) and binding affinity (Kd) at multiple template sites.
  • Utilized viral RNA templates designed to induce RT pausing due to secondary structures.

Main Results:

  • HIV-1 RTs exhibited significantly higher kpol values compared to HIV-2/SIV RTs across tested template sites.
  • No significant differences were observed in the Kd values between HIV-1 and HIV-2/SIV RTs.
  • HIV-1 RTs demonstrated more efficient DNA synthesis through pause sites, especially at low dNTP concentrations.

Conclusions:

  • HIV-1 RT's faster polymerization step (kpol) is a key adaptation to counteract low dNTP levels in non-dividing macrophages.
  • This enhanced kinetic property allows HIV-1 to maintain efficient reverse transcription despite cellular dNTP depletion.
  • The findings highlight distinct evolutionary strategies of lentiviruses for replication in specific host cell environments.