Nucleic Acids Research, Vol 27, Issue 16 3300-3309, Copyright © 1999 by Oxford University Press
T Hartshorne and W Toyofuku
Early pre-rRNA processing events were examined in the ancient protozoan
parasite Trypanosoma brucei and found to have both distinctive and
conserved features. Two 5'-ETS cleavages occur: A' and the newly discovered
A0. A' and A0 appear related to vertebrate and yeast primary pre-RNA
cleavage sites, respectively. However, trypanosomatid primary rRNA
transcripts can first be processed at the ITS1/5.8S boundary and 5'-ETS
sequences then removed by consecutive cleavages at A', A0 and A1 at the
5'-ETS/SSU rRNA junction. 5'-ETS sequences previously crosslinked to U3
snoRNA were tested for their roles in rRNA processing using our new tagged
rRNA system. Two distinct A'-adjacent sequence elements, which may pair
with U3 hinge bases, were specifically required for SSU rRNA production, as
was a downstream element. The latter element appears conserved with the
yeast 5'-ETS U3 binding sequence, required for A0, A1 and A2 cleavages, in
that they both share 10 bases complementary with U3 hinge sequences and lie
upstream from A0 and A1 sites located in a potential stem-loop structure.
The distinctive positioning of putative trypanosomatid U3 binding sites
with respect to A" and A0 cleavages suggests that different U3- dependent
mechanisms may direct each processing event.
ARTICLES
Two 5'-ETS regions implicated in interactions with U3 snoRNA are required for small subunit rRNA maturation in Trypanosoma brucei
Department of Biochemistry and Molecular Biology A-10, Albany Medical College, 47 New Scotland Avenue, Albany, NY 12208, USA. toinette_hartshorne@ccgateway.amc.edu
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