TRANSLATIONAL REPRESSION OF EF-1-ALPHA MESSENGER-RNA INVITRO

In this report we show that when 10000 X g supernatant extracts of growth arrested murine erythroleukemia (MEL) cells are incubated there is a rapid conversion of essentially all mRNAs to non-translating messenger ribonucleoprotein (RNP) particles. Most of these RNPs are readily translated in an initiation-dependent manner when added to a nuclease-treated rabbit reticulocyte lysate. A notable exception is the RNP containing eucaryotic elongation factor 1alpha (EF-1alpha) mRNA. The mRNA for poly(A)-binding protein behaved similarly to EF-1alpha. Previous work has demonstrated that the translation of both these mRNAs are repressed in vivo when the growth of a number of different mammalian cells is arrested [Slobin L. I. and Jordan, P. (1984) Eur J. Biochem. 145, 1984; Thomas, G. and Thomas, G. (1986) J. Cell Biol. 103, 19861. Translational activity of EF-1alpha mRNA could be restored by treating RNP particles with 0.5 M KCl, provided that the RNPs were separated from salt wash by chromatography on oligo(dT)-cellulose. Addition of the salt wash to total MEL cell mRNA significantly and selectively inhibited EF-1alpha mRNA translation, suggesting that a component of the salt wash acts as a trans-acting translational repressor of EF-1alpha mRNA.