SYNTHESIS AND CHARACTERIZATION OF A DNA COMPLEMENTARY TO PRE-UTEROGLOBIN MESSENGER-RNA

Uteroglobin, a progesterone‐induced uterine protein of the rabbit, is synthesized in cell‐free systems as a precursor containing 21 additional amino‐acids at its N‐terminal end. The mRNA for pre‐uteroglobin has been purified from the membrane‐bound polysomes of induced endometrium and used as template for the synthesis of a full copy complementary DNA. Final purification of the cDNA was based on hybridization to the template mRNA up to a low value of r0t(0.01 M · s) and digestion of the non‐hybridized cDNA by S1 nuclease. A comparison of the hybridization kinetics of the pre‐uteroglobin cDNA and rabbit globin cDNA to their respective templates indicates a nucleotide sequence complexity of 650 for pre‐uteroglobin mRNA, in agreement with the values obtained by sucrose gradient centrifugation and polyacrylamide gel electrophoresis in formamide. The melting temperature of the hybrids of pre‐uteroglobin cDNA to its template reflects the absence of mismatched sequences. This cDNA has been used to quantify pre‐uteroglobin mRNA sequences in the endometrial RNA from control animals and from animals treated sequentially with estradiol and progesterone. In agreement with the induction of uteroglobin‐synthesizing activity, there is a dramatic increase in the uterine content of pre‐uteroglobin mRNA after hormonal treatment. Part of this effect can be accounted for by hormonally induced cell proliferation. When expressed on a DNA basis there is a 50–100‐fold increase in the cellular content of pre‐uteroglobin mRNA following hormonal treatment. Copyright © 1979, Wiley Blackwell. All rights reserved