Nucleic Acids Research Advance Access originally published online on February 14, 2007
Nucleic Acids Research 2007 35(5):1612-1623; doi:10.1093/nar/gkm031
Nucleic Acids Research, 2007, Vol. 35, No. 5 1612-1623
© 2007 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.
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Group II intron in Bacillus cereus has an unusual 3' extension and splices 56 nucleotides downstream of the predicted site
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 July 21, 2006. Revised December 20, 2006. Accepted January 5, 2007.
All group II introns known to date fold into six functional domains. However, we recently identified an intron in Bacillus cereus ATCC 10987, B.c.I4, that splices 56 nt downstream of the expected 3' splice site in vivo (Tourasse et al. 2005, J. Bacteriol., 187, 5437–5451). In this study, we confirmed by ribonuclease protection assay that the 56-bp segment is part of the intron RNA molecule, and computational prediction suggests that it might form a stable stem-loop structure downstream of domain VI. The splicing of B.c.I4 was further investigated both in vivo and in vitro. Lariat formation proceeded primarily by branching at the ordinary bulged adenosine in domain VI without affecting the fidelity of splicing. In addition, the splicing efficiency of the wild-type intron was better than that of a mutant construct deleted of the 56-bp 3' extension. These results indicate that the intron has apparently adapted to the extra segment, possibly through conformational adjustments. The extraordinary group II intron B.c.I4 harboring an unprecedented extra 3' segment constitutes a dramatic example of the flexibility and adaptability of group II introns.
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