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Nucleic Acids Research, 1988, Vol. 16, No. 8 3497-3510
© 1988


Articles

Regulation of in vitro translation by double-stranded RNA in mammalian cell mRNA preparations

Gerg Pratt, Angela Galpine, Nigel Sharp, Susan Palmer and Michael J. Clemens

Department of Biochemistry, St George's Hospital Medical School Cranmer Terrace, London SW17 0RE, UK

Received January 11, 1988. Revised February 25, 1988. Accepted February 25, 1988.

Polyadenylated mRNA has been purified from a variety of human and mouse cell sources. These preparations are actively translated in the wheat germ cell-free system but have only poor ability to stimulate the nuclease-treated reticulocyte lysate. The translation of endogenous and exogenous globin mRNA is strongly inhibited by the poly(A)+ RNA preparations in reticulocyte lysates. Both polysomal and non-polysomal RNA have similar effects but poly(A)+ RNA is alaost 2000-fold nore inhibitory than poly(A)RNA on a weight basis. The inhibition is abolished in the presence a high concentration of poly(I).poly(C). Analysis of endogenous eIF-2 in the lysate reveals that the subunit becomes extensively phosphorylated in the presence of the inhibitory poly(A)+ RNA. Prolonged incubation of lysate with poly(A)+ RNA also causes some nucleolytic degradation of polysomal globin mRNA. These characteristics suggest that some eukaryotic cell mRNAs contain regions of double-stranded structure which are sufficiently extensive to activate translational control mechanisms in the reticulocyte lysate.


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