Selective cleavage by trypsin of the capsid protein VP1 of type 3 poliovirus results in improved sorting of cell bound virions

Summary

A large proportion of host cell-bound virions of poliovirus type 1 strain Mahoney (PV1/M) is known to elute to the culture medium during incubation at 37 °C, and only a fraction of the virions remaining cell-associated will successfully uncoat and contribute to the new replication cycle. We found that while the proportion of inoculum type 3 poliovirus strain Saukett (PV3/S) bound to GMK cells was of the same order as that of PV1/M, the bound PV3/S virions uncoated much less efficiently, as judged by velocity sedimentation analysis of virion disintegration. Rather, the majority of the cell-associated PV3/S viruses remained apparently unaffected for several hours within an unidentified intracellular compartment. Incubation of PV3/S with intestinal trypsin is known to result in selective cleavage of the capsid protein VP1 and striking antigenic changes. Trypsin treatment of stock PV3/S preparations did not affect the infectivity titre or modify single-cycle progeny virus yields significantly. However, the fate of the cell-bound inoculum virus was profoundly altered. Trypsin-treated PV3/S virions (PV3/S-Try) attached to GMK cells less tightly than the untreated PV3/S virus or PV1/M, and a relatively larger proportion of the cell-bound virus eluted to the medium during subsequent incubation at 36 °C. However, the fraction of virions remaining cell-associated rapidly disintegrated suggesting efficient uncoating. In accordance with these observations, one step growth curves of PV3/S-Try in all cell lines tested showed lowered eclipse phase titres compared to those obtained with the untreated PV3/S inocula. Similar effects were also demonstrated for type 3 poliovirus strain Sabin while trypsin-sensitive strains of the other two serotypes of poliovirus remained unaffected in this sense. The putative biological significance of the altered sorting of cell-bound PV3/S-Try virions is not known. It might be related to the observations that sensitivity of type 3 poliovirus strains to trypsin is conserved in spite of the fact that the target site of trypsin action is flanked by highly variable motives in an immunodominant antigenic site.