BIOCHEMICAL AND GENETIC STUDIES OF MULV ENV PROTEINS

Project Details

Description

The murine retroviruses are of interest both because of the pathology of
the diseases which they induce, and because of their utility as general
models of enveloped viruses. Many biologically important functions of
these viruses reside in the viral env gene products. The overall
objectives of our study are to determine structural-functional correlations
for the env proteins of different classes of MuLV. We have to date
developed procedures for isolating fragments of gp70 and p15(E) which
correspond to specific structural domains of these proteins. More detailed
studies of the primary, secondary, and tertiary structure of these domains
will be performed, including analyses of the oligosaccharide constituents
at specific glycosylation sites, localization of disulfide bonds, and
studies of the topographical organization of these domains in single
molecules and in oligomeric complexes of the env proteins. The
contribution of the individual domains to such functional properties of the
env proteins as receptor binding, cell fusion, and sensitivity to serum
neutralizing factor, will be determined both by direct studies with the
isolated subunits, and indirectly with antibodies directed against specific
domains. The function of various structural regions and features of the
env gene products in the processing, assembly, and biological activities of
these proteins will be analyzed by in vitro site-specific mutagenesis of
cloned viral DNAs, using synthetic oligonucleotides to introduce precise
amino acid changes into selected regions of the env gene products. Cells
expressing the mutant viruses will be isolated by cotransfection with a
selectable gene, and the structural, immunological, and functional
properties of the mutated gene products will be determined. In this way,
the functional roles of hydrophobic leader and transmembrane sequences,
conserved glycosylation sites and cysteine residues, and proteolytic
cleavage sites will be determined.
StatusFinished
Effective start/end date7/1/8512/31/92

Funding

  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health

ASJC

  • Medicine(all)

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