Nucleic Acids Research, Vol 27, Issue 11 2271-2282, Copyright © 1999 by Oxford University Press
B Michot, N Joseph, S Mazan and JP Bachellerie
Mechanisms of ITS2 excision from pre-rRNA remain largely elusive. In
mammals, at least two endonucleolytic cleavages are involved, which result
in the transient accumulation of precursors to 5.8S rRNA termed 8S and 12S
RNAs. We have sequenced ITS2 in four new species of the Mus genus and
investigated its secondary structure using thermodynamic prediction and
comparative approach. Phylogenetic evidence supports an ITS2 folding
organized in four domains of secondary structure extending from a preserved
structural core. This folding is also largely conserved for the previously
available mammalian ITS2 sequences, rat and human, despite their extensive
sequence divergence relative to the Mus species. Conserved structural
features include the structural core, containing the 3' end of 8S pre-rRNA
within a single-stranded sequence, and a stem containing the 3' end of the
12S pre-rRNA species. A putative, phylogenetically preserved pseudoknot has
been detected 1 nt downstream from the 12S 3' end. Two long
complementarities have also been identified, in sequences conserved among
vertebrates, between the pre-rRNA 32S and the snoRNA (small nucleolar RNA)
U8 which is required for the excision of Xenopus ITS2. The first
complementarity involves the 5.8S-ITS2 junction and 13 nt at the 5' end of
U8, whereas the other one occurs between a mature 28S rRNA segment known to
be required for ITS2 excision and positions 15-25 of snoRNA U8. These two
potential interactions, in combination with ITS2 folding, could organize a
functional pocket containing three cleavage sites and key elements for
pre-rRNA processing, suggesting a chaperone role for the snoRNA U8.
ARTICLES
Evolutionarily conserved structural features in the ITS2 of mammalian pre-rRNAs and potential interactions with the snoRNA U8 detected by comparative analysis of new mouse sequences
Laboratoire de Biologie Moleculaire Eucaryote du C.N.R.S., Universite Paul Sabatier, 118 route de Narbonne, 31062 Toulouse Cedex, France. bmichot@ibcg.biotoul.fr
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