Assessment of the role of the central DNA flap in human immunodeficiency virus type 1 replication by using a single-cycle replication system

J Virol. 2004 Mar;78(6):3170-7. doi: 10.1128/jvi.78.6.3170-3177.2004.

Abstract

In this study, reverse transcriptase (RT)- and integrase (IN)-defective human immunodeficiency virus type 1 (HIV-1) was transcomplemented with Vpr-RT-IN fusion proteins to delineate pol sequences important for HIV-1 replication. Our results reveal that a 194-bp sequence encompassing the 3'end of the IN gene and containing the central DNA flap is necessary and sufficient for efficient HIV-1 single-cycle replication in dividing and nondividing cells. Furthermore, we show that the central DNA flap enhances HIV-1 single-round replication by five- to sevenfold, primarily by facilitating nuclear import of proviral DNA. In agreement with previous reports, our data support a functional role of the central DNA flap during the early stages of HIV-1 infection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Nucleus / virology
  • DNA, Viral / biosynthesis*
  • DNA, Viral / chemistry
  • Gene Products, pol / chemistry
  • Gene Products, pol / genetics
  • Gene Products, vpr / genetics
  • Genetic Complementation Test
  • HIV Integrase / genetics
  • HIV Integrase / metabolism
  • HIV Reverse Transcriptase / genetics
  • HIV Reverse Transcriptase / metabolism
  • HIV-1 / genetics
  • HIV-1 / pathogenicity*
  • HIV-1 / physiology
  • Humans
  • Leukocytes, Mononuclear / virology
  • Recombinant Fusion Proteins
  • Virus Integration
  • Virus Replication
  • vpr Gene Products, Human Immunodeficiency Virus

Substances

  • DNA, Viral
  • Gene Products, pol
  • Gene Products, vpr
  • Recombinant Fusion Proteins
  • vpr Gene Products, Human Immunodeficiency Virus
  • HIV Integrase
  • HIV Reverse Transcriptase