ANALYSIS OF THE STRUCTURE AND BIOLOGICAL ACTIVITY OF PRUNUS NECROTIC RINGSPOT AND APPLE MOSAIC VIRUSES USING MONOCLONAL ANTIBODIES
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Date
1986-12
Type of Work
Department
Hood College Biology
Program
Hood College Biomedical Science
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Abstract
Monoclonal antibodies reactive with two ilarviruses, Prunus
necrotic ringspot virus (PNRSV) and apple mosaic virus (ApMV) were
utilized to determine the structural and biological properties of
the viruses. Three monoclonal antibodies (Mabs 1, 2 and 3) which
reacted only to ApMV in various ELISA tests were found to bind to
conformation independent epitopes which are hidden between or within
viral subunits. Three other monoclonal antibodies (Mabs 5, 6 and 7)
which reacted only to PNRSV in various ELISA tests were found to
bind to conformation dependent, externally located epitopes. One
monoclonal antibody, Mab 4, which reacted with both viruses was
found to bind to a conformation dependent, partialiy hidden epitope
on ApMV, but a conformation independent, external ,Titope on PNRSV.
Further, the three ApMV-specific antibodies reacted with viral coat
protein in Western blots. None of the three PNRSV-specific
antibodies reacted with viral coat protein in Western blots. Mab 4,
however, did react with PNRSV coat protein but not ApMV coat protein
in Western blots. Fragments of PNRSV coat protein were generated by
proteolysis, and, in Western blots, all were found to be bound by
Mab 4. Thus, attempts to isolate a specific polypeptide containing
the epitope were unsuccessful. In neutralization of infectivity
studies, Mab 4 blocked PNRSV infectivity although the antibody did
not precipitate the virus in Ouchterlony double diffusion tests.
This suggests that Mab 4 may bind to an epitope located in a region
of the coat protein necessary for infection. Although not yet
examined for PNRSV, a characteristic of other ilarviruses is coat
protein dependent initiation of infection, where coat protein, or a
subgenomic RNA which codes for coat protein, is required for the
RNAs to be infectious. Thus, Mab 4 may be a useful probe for
studying this process in PNRSV and other ilarviruses.