ABSTRACT
HeLa cells were made strictly dependent upon polyamine by growth in the presence of alpha-difluoromethylornithine, a specific inhibitor of ornithine decarboxylase. Under these conditions, the specific activity of the cellular polyamine pools eventually equilibrated to that of exogenously supplied [14C]putrescine; however, the process was very slow, requiring half-equilibration times of about 16 h for spermidine and 28 for spermine. Thus, the distribution of radioactivity in individual polyamines became a valid measure of polyamine content only after a continuous 4-day incorporation period. When propagated in polyamine-labeled cells, two picornaviruses were found to incorporate substantially different amounts of polyamine: about 0.6% of the cell pool for human rhinovirus 14 but only 0.04% for poliovirus. This content of polyamine was sufficient to neutralize nearly 27% of the negative charge of the RNA in human rhinovirus 14 but only 1.6% in poliovirus.
Subject(s)
Poliovirus/metabolism , Putrescine/metabolism , Rhinovirus/metabolism , Spermidine/metabolism , Spermine/metabolism , Eflornithine , HeLa Cells/metabolism , Humans , Kinetics , Ornithine/analogs & derivatives , Ornithine/pharmacology , Ornithine Decarboxylase InhibitorsABSTRACT
Crystals of rhinovirus 14 have been grown reproducibly. They diffract x-rays to a resolution of at least 3.5 A. The orthorhombic crystal unit cell contains two virions, each situated on a crystallographic twofold axis. At less than 30-A resolution, the space group approximates to 1222 with the particles possessing 222 pseudo crystallographic symmetry. The crystals are "isomorphous" with type I polio crystals [Finch, J. T. & Klug, A. (1959) Nature (London) 183, 1709-1714; Hogle, J. M. (1982) J. Mol. Biol. 160, 663-668], suggesting some similarities of structure between enteroviruses and rhinoviruses.
Subject(s)
Rhinovirus/ultrastructure , Crystallization , Poliovirus/ultrastructure , X-Ray DiffractionABSTRACT
Brain preparations from animals with scrapie or transmissible mink encephalopathy were phenol extracted and examined for the presence of pathogenic nucleic acid. Animals inoculated with various extracts remained healthy, and analysis on 2.6 to 5% polyacrylamide gels failed to detect a difference in extractable RNA species between infected and normal mink brain.
