ELECTROPHORETIC STUDY OF THE INTERACTION OF RADIOACTIVE POLIOVIRUS WITH COMPONENTS OF CULTURED CELLS

Abstract

Thorne , H. V. (University of Minnesota, Minneapolis). Electrophoretic study of the interaction of radioactive poliovirus with components of cultured cells. J. Bacteriol. 85: 1247–1255. 1963.—The interaction of P 32 -labeled poliovirus with the postnuclear fraction of HeLa cell homogenates was first examined using radioactivity to quantitate the reaction. The effects of virus and debris concentration, suspending medium, temperature, and time on the reaction were determined. Binding was independent of salt concentration up to 1.0 m and unaffected by calcium and magnesium ions at 10 −3 m . The postnuclear particulate components of homogenates of mammalian cells in culture were examined by sucrose gradient zone electrophoresis at pH 7.2 with a simple apparatus which permitted several simultaneous analyses. The distributions of components for different cell types were distinct and appeared to be influenced by conditions of culture. Addition of radioactive poliovirus to the homogenate before analysis was used to identify components with virus-binding activity. Activity of HeLa and human esophageal epithelium cell homogenates was found mainly in membranous fractions of relatively low electrophoretic mobility. Components with a broader spread of mobility were moderately active in cultured human amnion, but uncultured amnion had almost undetectable virus-binding activity and a distinct distribution of components. Rat heart cells and L cells did not bind poliovirus, but binding components were present in both ERK-1 and CRE rabbit lines.