Isolation and characterization of cytoplasmic inclusions from influenza A virus-infected cells

Abstract

Influenza A viruses induce the accumulation of electron-dense inclusions in the cytoplasm of infected cells during the latter stages of the replication cycle. Cell fractionation studies showed that these inclusions could be recovered in subcellular fractions containing ribosomes and polysomes. Isolation of these inclusions was accomplished by procedures involving RNase treatment of these fractions followed by repurification, or by fluorocarbon extraction and gradient centrifugation. Electron microscopy indicated that the isolated inclusions exhibited a major periodicity of approximately 8 nm with minor periodicities of approximately 4 nm. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the influenza virus coded nonstructural protein was the only protein component present in isolated inclusions.