G. Grafi et G. Galili, INDUCTION OF CYTOPLASMIC FACTORS THAT BIND TO THE 3' AU-RICH REGION OF HUMAN INTERFERON-BETA MESSENGER-RNA DURING EARLY DEVELOPMENT OF XENOPUS-LAEVIS, FEBS letters, 336(3), 1993, pp. 403-407
Certain endogenous Xenopus mRNAs, carrying a destabilizing 3' AU-rich
sequence, are unusually very stable in oocytes and become unstable onl
y after fertilization. In addition, heterologous short lived mRNA, con
taining 3' AU-rich sequences, appear to be very stable when injected i
nto Xenopus oocytes. In the present study, a human interferon beta (hu
-IFN beta) mRNA, carrying the destabilizing 3' AU-rich element, was us
ed as a probe to identify Xenopus proteins that specifically bind to t
he 3' AU-rich element as well as to study their relative levels during
early embryonic development. While three major proteins that specific
ally bind to the 3' AU-rich element were detected in human SV80 cells,
that naturally express hu-IFN beta (proteins termed AU-F1, F2 and F3)
, only two proteins, migrating similarly to the SV80 AU-F1 and AU-F3,
were detected in cytoplasmic extracts from Xenopus oocytes or eggs. Fo
llowing fertilization, the intensity of the Xenopus AU-F1 and AU-F3 pr
oteins increased considerably and a new protein, corresponding to SV80
AU-F2, was also detected. Cyclohexamide applied either at the morula
or at the early blastula stages reduced the intensity of the AU-bindin
g factors, while actinomycin D did not, indicating that the levels of
these factors during these stages are regulated posttranscriptionally.
In contrast, application of each of these metabolic inhibitors at the
late blastula stage increased the intensity of the AU-binding protein
s. The possible function of these AU-binding factors in regulating the
expression and half life of AU-rich mRNAs is discussed.