Nucleic Acids Research, Vol 27, Issue 24 4751-4758, Copyright © 1999 by Oxford University Press
K Duvel and GH Braus
Cleavage and polyadenylation of eukaryotic mRNA requires efficiency and
positioning elements in the 3'-untranslated region (3'-UTR) of the mRNA.
Specific point mutations were introduced into the yeast GCN4 3'- UTR to
detect sequence motifs which are involved in the positioning of the poly(A)
site. 3'-End proces-sing activities of different GCN4 3'- UTR alleles were
measured in an in vivo test system. Point mutations in an AAGAA motif
defocussed selection of the poly(A) sites of the GCN4 3'- UTR to various
additional poly(A) sites instead of the single site of the wild-type GCN4
3'-UTR. A strain with an intact wild-type GCN4 3'- UTR but impaired in
RNA15 encoding an RNA-binding processing factor showed a similar defocussed
pattern of poly(A) site selection. Remarkably, two additional sequence
motifs upstream of the AAGAA motif which resemble yeast efficiency motifs
independently affected poly(A) site positioning but not efficiency of
3'-end processing. Mutations in one motif resulted in an additional
upstream poly(A) site. Alterations of the other motif shifted the poly(A)
sites exclusively to two downstream poly(A) sites. These data suggest
several contact points between the precursor mRNA and the polyadenylation
machinery in yeast.
ARTICLES
Different positioning elements select poly(A) sites at the 3'-end of GCN4 mRNA in the yeast Saccharomyces cerevisiae
Institute of Microbiology, Georg-August-University, Grisebachstrasse 8, D-37077 Gottingen, Germany.
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