HIV 1 INTEGRATION

Project: Research project

Description

DESCRIPTION: The goals of this project
are to examine the integration reaction, in which human immunodeficiency
virus (HIV) DNA is incorporated into cellular DNA, and to understand the
role of integration in the pathogenesis of AIDS. Initial experiments
proposed are to characterize integration in blood cells of HIV-infected
individuals. Polymerase chain reaction (PCR) will be performed to measure
the levels of integrated and unintegrated HIV DNA, the compartmentalization
of HIV DNA between cytoplasm and nuclease, and the forms of unintegrated
HIV DNA. The investigator anticipates that these data will localize the
probable site in the integration pathway of any observed inefficiency of
integration. The influence of cell type on integration will also be
examined. The remaining sections of this application describes a combined
genetic and biochemical approach that focuses on HIV integrase (IN).
Infections with virions containing mutations at defined positions in the 3'
region of the HIV pol (specifies viral reverse transcriptase) gene will be
examined for specific defects in integration. Quantitative analysis will
indicate whether integration is required for productive HIV infection. To
begin to define domains of IN, mutated IN proteins will be compared with
wild-type (wt) proteins for activity in assay systems that explore each
step in the integration pathway, including DNA binding, endonuclease,
covalent linkage and DNA joining. These assays will also be used to
compare purified integrases from viruses with different pathogenic
potential. The unique aspect of this plan involves expressing and
purifying IN from visna virus, a lentivirus that is capable of productive
infection without integration. Analysis of visna virus IN may provide
insights into HIV IN which may be relatively inefficient at catalyzing
integration.
StatusFinished
Effective start/end date8/1/915/31/97

Funding

  • National Institutes of Health: $107,634.00
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health

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Integrases
DNA
Visna-maedi virus
Lentivirus
Deoxyribonuclease I
RNA-Directed DNA Polymerase
Infection
Virion
Blood Cells
Cytoplasm
Acquired Immunodeficiency Syndrome
Proteins
Research Personnel
Viruses
Polymerase Chain Reaction
Mutation
Genes