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

Nucleotide-induced stable complex formation by HIV-1 reverse transcriptase

W Tong1, C D Lu, S K Sharma

  • 1Department of Biochemistry and Molecular Biology, University of Miami School of Medicine, Florida 33101, USA.

Biochemistry
|May 13, 1997
PubMed
Summary
This summary is machine-generated.

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

[Analysis of strong invasive clinical features and poor prognosis in macrotrabecular-massive hepatocellular carcinoma].

Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology·2024
Same author

[Impact of regional positive lymph node ratio of gallbladder carcinoma on prognosis].

Zhonghua zhong liu za zhi [Chinese journal of oncology]·2022
Same author

[Efficacy of in-situ full-left/full-right split liver transplantation for adult recipients using the living donor liver transplantation technique:a single-center report of 25 cases].

Zhonghua wai ke za zhi [Chinese journal of surgery]·2022
Same author

[Evaluations of total hilar en bloc resection and reconstruction in comparison with portal vein resection and reconstruction in treatment of perihilar cholangiocarcinoma].

Zhonghua wai ke za zhi [Chinese journal of surgery]·2021
Same author

Delta Helicase Corresponds to KIAA0221: A Link Between DNA Polymerase Delta and RNA Surveillance Complex.

TheScientificWorldJournal·2018
Same author

[PAHs exposure and occupational health risk assessment of workers in coal tar pitch factory].

Zhonghua lao dong wei sheng zhi ye bing za zhi = Zhonghua laodong weisheng zhiyebing zazhi = Chinese journal of industrial hygiene and occupational diseases·2018
Same journal

Aromatic Cage-Directed Azide-Methyllysine Photochemistry for Profiling Nonhistone Interacting Partners of the MeCP2 Methyl-CpG-Binding Domain.

Biochemistry·2026
Same journal

Differential Hydroxypyruvate Processing by <i>E. coli</i> and <i>P. aeruginosa</i> DXP Synthases Reveals Preferential Xylulose 5-Phosphate Formation by the <i>P. aeruginosa</i> Enzyme.

Biochemistry·2026
Same journal

Structural and Functional Characterization of Heterologous Nitrogenase Complexes.

Biochemistry·2026
Same journal

Discovery of Bacterial Unspecific Peroxygenases.

Biochemistry·2026
Same journal

Lactate Biology: Subcellular Routing and Chemical Form Define Function.

Biochemistry·2026
Same journal

Nature's Anaerobic Toolkit: Glycyl Radical Enzymes and Their Expanding Functional and Mechanistic Diversity.

Biochemistry·2026
See all related articles

Human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT) undergoes conformational changes when binding nucleotides. HIV-1 RT discriminates against non-complementary nucleotides and certain analogues during DNA synthesis.

Area of Science:

  • Molecular Biology
  • Virology
  • Biochemistry

Background:

  • Reverse transcriptase (RT) is a crucial enzyme for human immunodeficiency virus type 1 (HIV-1) replication.
  • Understanding RT's substrate binding and conformational changes is key to developing antiviral therapies.
  • Previous studies have investigated HIV-1 RT's interaction with various nucleotide substrates.

Purpose of the Study:

  • To investigate the nucleotide substrate-induced conformational changes in HIV-1 RT.
  • To analyze the mechanism of discrimination between complementary and non-complementary deoxynucleotide triphosphates (dNTPs) and nucleotide analogues by HIV-1 RT.
  • To determine the optimal primer/template length for dead-end complex formation.

Main Methods:

  • Nondenaturing gel electrophoresis was employed to study the formation of dead-end complexes.

Related Experiment Videos

  • Dead-end complexes were formed between HIV-1 RT, a chain-terminated primer, and a DNA template.
  • The effect of complementary and non-complementary dNTPs, ribonucleotides, and nucleotide analogues on complex formation was assessed.

    • Main Results:

    • Dead-end complex formation was significantly more efficient with complementary dNTPs compared to non-complementary dNTPs or ribonucleotides (over 2000-fold less efficient).
    • The apparent dissociation constant (Kd) varied for different dTTP and dGTP analogues, indicating differential substrate discrimination.
    • Optimal primer/template duplex length for dead-end complex formation was 20-32 base pairs, with mismatches or specific chain terminators affecting complex stability.

    Conclusions:

    • HIV-1 RT exhibits distinct conformational changes upon binding complementary nucleotides, facilitating efficient DNA synthesis.
    • The enzyme effectively discriminates against non-complementary nucleotides and certain analogues, contributing to its fidelity.
    • These findings provide insights into the molecular mechanisms underlying nucleotide analogue discrimination by HIV-1 RT, relevant for antiviral drug design.