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Research
Interest
The
lifecycle of HIV is quite
complicated and requires several distinct steps including cell fusion,
reverse
transcription, integration, transcription and translation, viral
assembly,
budding and release, and maturation. HIV recruits the help of host
machinery
along the way from viral entry to viral assembly. During viral assembly
tRNALys,3
is packaged into the virion so it can be used as a primer for reverse
transcription later on. It has also been shown that this tRNA is
selectively
packaged with the help of the cellular protein lysyl-tRNA synthetase
(LysRS).
Other host cofactors have been shown to interact with HIV derived
proteins in
the cell, including the APOBEC protein, which in vif-negative viruses
blocks
budding and release of viral particles.
The
goal of this project is to
precisely map the protein-protein interactions of HIV proteins, such as
gag and gag-pol,
with host cell cofactors including LysRS and APOBEC. We
will use mass-spec footprinting to identify interaction sites by
chemical
modification of histidines, arginines, tryptophans, and tyrosines. In
general
the protein of interest will be chemically treated to modify one or
more of the
four amino acids listed above in the absence of interacting protein
partner and
mass-spec analysis will be run to identify regions that were modified.
A
parallel experiment in the presence of interacting protein partner will
be run
and mass-spec analysis will reveal if any previously modified regions
were
protected by close interactions with the counterpart protein. This
information
will be mapped back to the 3D structure of the individual proteins and
potential binding sites will be identified. This work will be carried
out in
collaboration with Dr. Mamuka Kvaratskhelia (OSU).
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