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Related Experiment Videos

Subcellular localization and integration activities of rous sarcoma virus reverse transcriptase.

Susanne Werner1, Patrick Hindmarsh, Markus Napirei

  • 1Max-Planck-Institut für Molekulare Physiologie, Abteilung Physikalische Biochemie, 44227 Dortmund, Germany.

Journal of Virology
|May 22, 2002
PubMed
Summary

Avian Rous sarcoma virus (RSV) reverse transcriptase (RT) subunits are found in the nucleus, suggesting a role in transporting viral DNA. However, RSV RT

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Area of Science:

  • Virology
  • Molecular Biology
  • Enzymology

Background:

  • Avian retroviral reverse transcriptases (RTs), like Rous sarcoma virus (RSV) RT, possess an integrase domain, unlike their non-avian counterparts.
  • RSV integrase contains a nuclear localization signal, crucial for integrating proviral DNA into the host genome.
  • Previous studies indicated that RSV completes proviral DNA synthesis within the host cell nucleus.

Purpose of the Study:

  • To investigate the subcellular localization of RSV RT.
  • To determine if the integrase domain of RSV RT possesses integration activity.
  • To elucidate the role of RSV RT in nuclear transport and viral DNA integration.

Main Methods:

  • Analyzing the subcellular localization of heterodimeric RSV RT (alphabeta) and individual subunits (alpha, beta) using expression systems.

Related Experiment Videos

  • Assessing the nicking and joining activities of heterodimeric RSV RT (alphabeta) using oligodeoxynucleotide and supF gene-based assays.
  • Evaluating the in vitro integration efficiency of RSV RT alphabeta compared to free RSV integrase.

    • Main Results:

    • The heterodimeric RSV RT (alphabeta) and the beta subunit localize to the nucleus, while the alpha subunit is primarily cytoplasmic.
    • RSV RT alphabeta demonstrates in vitro integration activity, albeit with significantly lower efficiency (30-fold less) than free RSV integrase.
    • Integration activity of RSV RT alphabeta can be enhanced by co-factors like HIV-1 nucleocapsid protein or HMG-I(Y).

    Conclusions:

    • RSV RT's nuclear localization suggests a role in transporting the preintegration complex into the nucleus.
    • RSV RT itself is unlikely to be involved in the in vivo integration process due to its low efficiency.
    • The integrase domain within RSV RT may have functions beyond direct integration, potentially in complex assembly or transport.