Translation and stability of human globin mRNA in Xenopus oocytes.

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Abstract

Human globin messenger RNA (mRNA) prepared from erythroid cells of patients with sickle cell anemia has been translated in Xenopus laevis oocytes. Addition of hemin to the injected mRNA causes total globin synthesis to increase and the ratio of alpha- to betas-globin synthesis (alpha/betas ratio) to approach unity. To determine the effect of the length of the poly-(A) segment on human globin mRNA stability, 10 S globin mRNA was fractionated into poly-(A)-poor fractions by oligo (dT)-cellulose column chromatography. When oocytes are injected with each of these fractions, translation of the poly-(A)-rich globin mRNA is sustained for a longer period than that of the poly-(A)-poor mRNA. Regardless of the mRNA fraction injected, the alpha/betas ratio of the synthesized globin decreases as the injected oocytes are incubated for longer periods. The results indicate that in frog oocytes poly-(A)-rich mRNA has greater translational stability than poly-(A)-poor mRNA, AND beta-mRNA has greater stability than alpha-mRNA with comparable poly-(A) content.

Authors

G M Maniatis, F Ramirez, A Cann, P A Marks, A Bank