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Nucleic Acids Research, 1978, Vol. 5, No. 12 4781-4794
© 1978

Articles

Identification of a large preculsor of vitellogenin mRNA in the liver of estradiol-treated chicks

Jean-Pierre Jost, Gundula Pehling, Takeshi Ohno and Peter Cozens

Friedrich Miescher-Institut P.O. Box 273, CH-4002 Basel, Switzerland

Received October 19, 1978. Studies were performed to determine whether vitellogenin mRNA from avian liver has a precursor molecule or not. Total cellular RNA was prepared from estradiol-treated chicken liver in the presence of 8 M guanidine HCl, 2-mercaptoethanol and aurintricarboxylic acid. After denaturation, RNA was fractionated on sodium dodecylsulfate-sucrose gradients and large size RNA was analyzed under stringent conditions on 85% formamide-sucrose gradients at 25°C. RNA fractions collected from the gradients were hybridized with vitellogenin (3H)-cDNA Besides mature vitellogenin mRNA (32S, 7,000 nucleotides) vitellogenin sequences were also found in RNA fractions ranging from 38–50S with a peak at 45–50S (12–15,000 nucleotides). Only 5–10% of the putative 38–50S pmRNA is polyadenylated. We calculated that the half-life of vitellogenin pmRNA is about 3–4 minutes. We conclude that vitellogenin mRNA has a precursor which is twice the size of the mature mRNA.


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