Nontranslated cellular mRNAs are associated with the cytoskeletal framework in influenza virus or adenovirus infected cells

Abstract

In an effort to understand the molecular mechanisms underlying the selective shutoff of host protein synthesis in influenza virus and adenovirus infected cells, we analyzed the subcellular location of representative cellular and viral mRNAs. Earlier work has shown that the majority of cellular mRNAs remain polysome associated after infection by either virus and that both the initiation and elongation steps of host protein synthesis were blocked in infected cells (M. G. Katze, D. DeCorato, and R. M. Krug, J. Virol., 60, 1027–1039, 1986). The present study was undertaken to test whether these cellular mRNAs were rendered nontranslatable during infection as a result of their dissociation from the cytoskeleton framework. HeLa cells were fractionated into subcellular components by first gently disrupting the cells with Triton X-100 yielding the soluble fraction (SOL); the cytoskeleton (CSK) fraction was obtained from the Triton insoluble material by the double detergent treatment of Tween-40 and sodium deoxycholate. In uninfected cells the majority of host mRNAs were associated with polysomes which were exclusively bound to the CSK as would be expected of actively translated mRNAs. The cellular mRNAs also remained almost totally associated with the cytoskeleton in adenovirus and influenza virus infected cells despite the fact these mRNAs are not translated during infection. Indeed, the host mRNAs and the efficiently translated viral mRNAs were CSK associated to the same extent. In contrast to the adenovirus and influenza systems, significant amounts of cellular mRNAs were dissociated from the CSK and found in the SOL fraction of poliovirus infected cells as others have reported. In accordance with the biochemical analysis, morphological studies utilizing electron microscopy revealed that the cytoskeleton remained relatively intact during adenovirus and influenza infection but was substantially reorganized in poliovirus infected cells. We conclude that translational regulatory events are likely different in the poliovirus system and that cytoskeletal association of mRNAs may be required but is not sufficient for efficient mRNA translation during adenovirus or influenza virus infection.