Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Fidelity of HIV-1 reverse transcriptase.

B D Preston1, B J Poiesz, L A Loeb

  • 1Department of Pathology, University of Washington, Seattle 98195.

Science (New York, N.Y.)
|November 25, 1988
PubMed
Summary

The human immunodeficiency virus type 1 (HIV-1) reverse transcriptase makes frequent errors during DNA replication, contributing to the virus's rapid evolution. Understanding these errors may help develop new antiviral therapies.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

HESI/FDA workshop on immunomodulators and cancer risk assessment: Building blocks for a weight-of-evidence approach.

Regulatory toxicology and pharmacology : RTP·2016
Same author

Targeted delivery of anti-cancer growth inhibitory peptides derived from human alpha-fetoprotein: review of an International Multi-Center Collaborative Study.

Journal of drug targeting·2010
Same author

Werner Syndrome, aging and cancer.

Genome dynamics·2008
Same author

Mutations in human DNA polymerase eta motif II alter bypass of DNA lesions.

The EMBO journal·2001
Same author

Transduction of cellular sequence by a human immunodeficiency virus type 1-derived vector.

Journal of virology·2001
Same author

In vivo mutagenesis by Escherichia coli DNA polymerase I. Ile(709) in motif A functions in base selection.

The Journal of biological chemistry·2001

Area of Science:

  • Molecular Biology
  • Virology
  • Biochemistry

Background:

  • Human immunodeficiency virus type 1 (HIV-1) exhibits significant genetic diversity and rapid evolution.
  • The reverse transcriptase enzyme is crucial for HIV-1 replication and is known to have a high error rate.

Purpose of the Study:

  • To quantify the fidelity of purified HIV-1 reverse transcriptase during DNA polymerization in vitro.
  • To investigate the specific types and frequencies of base-substitution errors introduced by HIV-1 reverse transcriptase.
  • To explore the implications of HIV-1 reverse transcriptase error rates for viral hypermutability and antiviral drug development.

Main Methods:

  • In vitro DNA polymerization assays using purified HIV-1 reverse transcriptase.
  • Analysis of base-substitution errors in bacteriophage phi X174 amber3 DNA.
  • Determination of misincorporation rates opposite template adenine residues.

Main Results:

  • HIV-1 reverse transcriptase introduced base-substitution errors at frequencies of 1/2000 to 1/4000.
  • The enzyme showed a specificity for misincorporation: A:C >> A:G > A:A.
  • An estimated five to ten errors per HIV-1 genome occur per replication cycle in vivo due to reverse transcriptase activity.

Conclusions:

  • The high in vitro error rate of HIV-1 reverse transcriptase likely contributes significantly to the hypermutability of the AIDS virus in vivo.
  • The observed specificity of misincorporation by HIV-1 reverse transcriptase may inform the rational design of antiviral nucleoside analogs.

Related Experiment Videos