Abstract
Triton-insoluble cytoskeletons were prepared from uninfected and adenovirus-infected KB cells. Gradient analysis showed that all cellular polyribosomes were present in the cytoskeletons. After disaggregation of the polyribosomes, in vivo or in vitro, most of the messenger RNA (mRNA) remained associated with the cytoskeletal framework. Translation experiments showed that most mRNA species were present in a bound (cytoskeletal), as well as in an unbound state. However, whereas some mRNA species were predominant as unbound mRNP particles, other mRNA species were almost exclusively found in polyribosomes associated with the cytoskeletal framework. Incubation of cytoskeletons in an mRNA-dependent reticulocyte cell-free system revealed synthesis of the same set of polypeptides as took place when using whole cells. Furthermore, the gradual shift from translation of cellular to translation of viral mRNA species during late phase of productive infection with adenovirus could also be followed when cytoskeletons were translated in the cell-free system. These results support the hypothesis that Triton X-100 extraction does not remove actively translating mRNA from the cells, thus suggesting a functional relationship between mRNA translation and mRNA binding to a cytoskeletal framework.
Published Version
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