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Nature Structural & Molecular Biology
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U2–U6 RNA folding reveals a group II intron-like domain and a four-helix junction

Nature Structural & Molecular Biologyvolume 11, pages1237–1242 (2004)Cite this article

ACorrigendum to this article was published on 01 January 2005

Abstract

Intron removal in nuclear precursor mRNA is catalyzed through two transesterification reactions by a multi-megaDalton ribonucleoprotein machine called the spliceosome. A complex between U2 and U6 small nuclear RNAs is a core component of the spliceosome. Here we present an NMR structural analysis of a protein-free U2–U6 complex fromSaccharomyces cerevisiae. The observed folding of the U2–U6 complex is a four-helix junction, in which the catalytically important AGC triad base-pairs only within U6 and not with U2. The base-pairing of the AGC triad extends the U6 intramolecular stem-loop (U6 ISL), and the NMR structure of this extended U6 ISL reveals structural similarities with domain 5 of group II self-splicing introns. The observed conformation of the four-helix junction could be relevant to the first, but not the second, step of splicing and may help to position the U6 ISL adjacent to the 5′ splice site.

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Figure 1: Sequence of the U2–U6 complex fromSaccharomyces cerevisiae.
Figure 2: NMR evidence for secondary structure of U2–U6 reveals a four-helix junction conformation.
Figure 3: NMR structure of the extended U6 ISL, as observed in the context of the U2–U6 complex.
Figure 4: Structural comparison of the extended U6 ISL and D5.
Figure 5: Model for U2–U6 conformational change after first step of splicing.

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Acknowledgements

We thank C.J. McManus for sample preparation, and M. Tonelli and the National Magnetic Resonance Facility (NMRFAM) staff for technical support. NMR studies were carried out at NMRFAM with support from the US National Institutes of Health (NIH) Biomedical Technology Program and additional equipment funding from the University of Wisconsin, the US National Science Foundation (NSF) Academic Infrastructure Program, the NIH Shared Instrumentation Program, the NSF Biological Instrumentation Program, and the US Department of Agriculture. This investigation was supported by NIH grant GM65166 to S.E.B. and by NIH predoctoral training grant T32 GM007215 to D.G.S.

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Authors and Affiliations

  1. Department of Biochemistry, University of Wisconsin-Madison, 433 Babcock Drive, Madison, 53706, Wisconsin, USA

    Dipali G Sashital, Gabriel Cornilescu & Samuel E Butcher

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  1. Dipali G Sashital

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  2. Gabriel Cornilescu

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  3. Samuel E Butcher

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Correspondence toSamuel E Butcher.

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Supplementary information

Supplementary Fig. 1

U2-U6 complexes used in NMR study. (PDF 78 kb)

Supplementary Fig. 2

NMR evidence for four-helix junction in all U2–U6 complexes. (PDF 126 kb)

Supplementary Fig. 3

Overlay of 2D NOESY sequential region for extended U6 ISL and U2–U6 complex. (PDF 123 kb)

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Sashital, D., Cornilescu, G. & Butcher, S. U2–U6 RNA folding reveals a group II intron-like domain and a four-helix junction.Nat Struct Mol Biol11, 1237–1242 (2004). https://doi.org/10.1038/nsmb863

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