Characteristics of poly(A)-degrading factor present in the avian oocytes and early embryos.

Abstract

The presence of poly(A)-degrading activity was studied in vitro in the quail and mouse oocytes and early embryos using 3H-poly(A) as a substrate. The activity was measured by adsorption of the undegraded substrate to DE-81 filter paper discs, by chromatographic separation on Sephadex G-50 column and by agarose gel electrophoresis followed by transfer onto a Zeta-probe membrane (BioRad, Richmond, CA) and autoradiography. High poly(A)-degrading activity was found in the quail previtellogenic and vitellogenic oocytes and lower activity in the early embryos from cleavage stage to gastrulation. This activity is localized predominantly in the nucleus and, to a lesser degree, in the cytoplasm and in the vitellus of vitellogenic oocytes. The length of the poly(A) degradation product was estimated to be of about (A)10. Optimum activity was at pH 8.7 and at Mn2+ concentration of 0.5 mM. This makes the deadenylating enzyme from the quail oocytes similar to endoribonuclease IV from the chick and quail oviducts (Müller [1976] Eur. J. Biochem., 70:241-248; Müller [1976], Eur. J. Biochem., 70:249-258). We suggest that the poly(A)-degrading enzyme, similar to endoribonuclease IV found in the quail oocytes, might be the "deadenylating factor" reported in Xenopus oocytes (Varnum et al. [1992] Dev. Biol., 153:283-290). Such poly(A)-degrading activity is undetectable in unfertilized mouse eggs; however, a slight, statistically insignificant tendency for poly(A) degradation was seen in two-cell embryos.