REPLICATION OF VESICULAR STOMATITIS VIRUS

Project Details

Description

The aims of this project are to study the macromolecular regulatory
steps during the replication of vesicular stomatitis virus (VSV)
and to discover underlying mechanisms by which viruses cause
disease, with the hope that novel counter measures will be
discovered for the better control of viral diseases. Molecular
biochemical techniques will be used throughout. This will be
coupled with genetic and biological approaches. The glycoprotein G of VSV will be studied in relation to its
natural cleavage products Gs and anchor. Their functions in vitro
and as immunogens in mice will be delineated. Virions, containing
only anchors, will be tested for immune enhancement using non-
neutralizing antibodies and macrophages. The point of cleavage and
the overall structure of Gs will be determined by terminal amino
acid analysis and, eventually, X-ray crystallography. Also, how
efficiently VSV donates its G to HIV during coinfection of H9 cells
with RF-HIV will be assessed. Pseudotypes of HIV (VSV) will be
characterized by their susceptibility to specific neutralizing
antibodies and by their expanded host range. The role of defective interfering (DI) particles in eliciting or
modulating immunogenic responses will be studied in mice and in
reconstituted immune cell in vitro. Preliminary data show that DI
particles differ from standard infectious VSV in their interactions
with T lymphocytes. VSV mRNA's will be utilized to detect cytoplasmic regulatory
controls. Cloned cDNA's will be engineered to contain mutations,
deletions or extra sequences. mRNA made from these cDNA's will be
tested in vitro or in vivo for translational efficiency, ribosome
binding and dissociation, mRNA stability, and targeting to membrane
organelles. Initially, engineered sequences will be made in the
5' and 3' non-coding leader regions Then nonessential coding
regions and regions containing other open reading frames will be
altered. If sequences in the mRNA are found to respond to
cytoplasmic regulation then those sequences will be used in binding
assays to detect the proteins or ribonucleoproteins that may be
responsible. Also, peptides made from these minor open reading
frames will be used to make antibody for the purpose of determining
whether these products exist during vsv infections.
StatusFinished
Effective start/end date12/31/891/31/95

Funding

  • National Institute of Allergy and Infectious Diseases
  • National Institute of Allergy and Infectious Diseases
  • National Institute of Allergy and Infectious Diseases
  • National Institute of Allergy and Infectious Diseases

ASJC

  • Medicine(all)
  • Immunology and Microbiology(all)
  • Virology
  • Immunology

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