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Nucleic Acids Research Advance Access originally published online on March 21, 2009
Nucleic Acids Research 2009 37(10):3202-3214; doi:10.1093/nar/gkp186
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Nucleic Acids Research, 2009, Vol. 37, No. 10 3202-3214
© 2009 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

RNA

A conserved 3' extension in unusual group II introns is important for efficient second-step splicing

Fredrik B. Stabell, Nicolas J. Tourasse and Anne-Brit Kolstø*

Laboratory for Microbial Dynamics (LaMDa), Department of Pharmaceutical Biosciences, University of Oslo, Oslo, Norway

*To whom correspondence should be addressed. Tel: +47 22 85 69 23; Fax: +47 22 84 49 44; Email: a.b.kolsto{at}farmasi.uio.no

Received November 19, 2008. Revised March 5, 2009. Accepted March 6, 2009.

The B.c.I4 group II intron from Bacillus cereus ATCC 10987 harbors an unusual 3' extension. Here, we report the discovery of four additional group II introns with a similar 3' extension in Bacillus thuringiensis kurstaki 4D1 that splice at analogous positions 53/56 nt downstream of domain VI in vivo. Phylogenetic analyses revealed that the introns are only 47–61% identical to each other. Strikingly, they do not form a single evolutionary lineage even though they belong to the same Bacterial B class. The extension of these introns is predicted to form a conserved two-stem–loop structure. Mutational analysis in vitro showed that the smaller stem S1 is not critical for self-splicing, whereas the larger stem S2 is important for efficient exon ligation and lariat release in presence of the extension. This study clearly demonstrates that previously reported B.c.I4 is not a single example of a specialized intron, but forms a new functional class with an unusual mode that ensures proper positioning of the 3' splice site.


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