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Nucleic Acids Research, 1989, Vol. 17, No. 22 9003-9014
© 1989

MOLECULAR BIOLOGY

Ribosome loading, but not protein synthesis is required for estrogen stabilization of Xenopus laevis vitellogenin mRNA

John E. Blume1 and David J. Shaprio

Department of Biochemistry, University of Illinois 1209 West California St., Urbana, IL 61801, USA

Received September 5, 1989. Revised October 11, 1989. Accepted October 11, 1989.

We have examined the effect of protein synthesis and of ribosome loading on the estrogen-mediated stabilization of hepatic Xenopus laevis vitellogenin mRNA. Removal of estradiol-17ß from the culture medium, which destabilizes vitellogenin mRNA, does not alter the density of ribosomes on polysomal vitellogenin mRNA, or change the proportion of vitellogenin mRNA associated with the endoplasmic reticulum. Cycloheximide, which inhibits elongation, without changing the density of ribosomes on vitellogenin mRNA, does not block estrogen-mediated stabilization. In contrast, 2-(4-methyl-2,6-dinitroanilino)-N-methylpropionamide, (MDMP), which inhibits initiation, greatly reduces the density of ribosomes on vitellogenin mRNA, and completely blocks estrogen-mediated stabilization. Vitellogenin mRNA in MDMP treated cells is degraded at a rate similar to that seen when untreated cells are transferred from medium containing estrogen to estrogen-free medium. This suggests that a ribosorne-associated degradative system may not be responsible for vitellogenin mRNA degradation. The failure of estrogen to stabilize vitellogenin mRNA in MDMP-treated cells is not due to the release of vitellogenin mRNA from the endoplasmic reticulum. Vitellogenin mRNA in MDMP-treated cells remains associated with the ezuioplasmic reticulum in small polysomes containing 3–5 ribosomes. These data demonstrate that maintaining a high density of ribosomes on vitellogenin mRNA, but not continuing protein synthesis, is necessary for estrogen-mediated stabilization of vitellogenin mRNA.

1 Present address: Department of Biochemistry, Roche Institute of Molecular Biology, Nutley, New Jersey.


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