Nucleic Acids Research, Vol 27, Issue 2 429-438, Copyright © 1999 by Oxford University Press
C Phillips, CB Kyriakopoulou and A Virtanen
We have previously shown that a distal GU-rich downstream element of the
mouse IgM secretory poly(A) site is important for polyadenylation in vivo
and for polyadenylation specific complex formation in vitro. This element
can be predicted to form a stem-loop structure with two asymmetric internal
loops. As stem-loop structures commonly define protein RNA binding sites,
we have probed the biological activity of the secondary structure of this
element. We show that mutations affecting the stem of the structure abolish
the biological activity of this element in vivo and in vitro at the level
of cleavage and polyadenylation specificity factor/cleavage stimulation
factor complex formation and that both internal loops contribute to the
enhancing effect of the sequence in vivo. Lead (II) cleavage patterns and
RNase H probing of the sequence element in vitro are consistent with the
predicted secondary structure. Furthermore, mobility on native PAGE
suggests a bent structure. We propose that the secondary structure of this
downstream element optimizes its interaction with components of the
polyadenylation complex.
ARTICLES
Identification of a stem-loop structure important for polyadenylation at the murine IgM secretory poly(A) site
Department of Genetics and Pathology, Uppsala University, Box 589, SE- 751 23 Uppsala, Sweden.
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