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Although a vaccine exists for Measles, a highly communicable, respiratory disease, the infective virus (measles virus, MV) remains a lethal pathogen.
Incomplete compliance with vaccination in the developed world and low vaccination coverage in developing countries results in outbreaks, making
the development of MV inhibitors desirable. This study characterizes the mechanism of antiviral activity and identifies the molecular target of a
non-nucleoside inhibitor family of the MV RNA-dependent RNA polymerase (RdRp) complex which we have developed.
The goals of this project were
1. to characterize a highly potent analog of an HTS hit antiviral,
2. to generate resistant virions through directed evolution and assess robustness of resistance, and
3. to identify the molecular basis for resistance through RT-PCR and DNA sequencing.
After quantification of the compound’s inhibitory effect against both strains, growth curves were generated to assess each strain’s fitness in vitro.
Gene sequencing was used to identify mutations in the viral genome. Our study indicates that compound resistance does not substantially reduce
viral fitness in vitro. Mutations were found only in the viral polymerase (L) protein, and the L protein is believed to be the compound’s physical target.
This conclusion will be tested through reengineering of mutations in a cDNA copy of the viral genome and followed by recovery of recombinant virions
and assessment of their sensitivity to the compound.
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