Use of drug-resistance mutants to identify functional regions in picornavirus capsid proteins
The WIN drugs and similar hydrophobic compounds that insert into the capsid of picornaviruses have been shown to block viral uncoating. In some of the human rhinoviruses they also block attachment of virus to cells. Spontaneously occurring drug-resistant mutants of human rhinovirus 14 and poliovirus type 3 were selected for their ability to make plaques in the presence of the selecting drug. The HRV-14 mutants either prevented drug binding or allowed the virus to attach to cells in the presence of drug. About two thirds of the poliovirus mutants were dependent on the presence of drug for plaque formation. In single cycle growth curves, drug was not required for the formation of drug-dependent progeny virus. However, progeny virus grown without drug never accumulated outside of cells, thus making the formation of plaques impossible. This behavior was apparently caused by the extreme thermolability of these mutants. In the absence of drug, heating to 37°C rapidly converted them to non-infectious particles with a sedimentation coefficient of 135S.
KeywordsPlaque Assay Wild Type Virus Progeny Virus Human Rhinovirus 135S Particle
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