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Autocatalytic RNA cleavage in the human β-globin pre-mRNA promotes transcription termination

Nature volume 432, pages 526–530 (2004)Cite this article

Abstract

New evidence indicates that termination of transcription is an important regulatory step, closely related to transcriptional interference1 and even transcriptional initiation2. However, how this occurs is poorly understood. Recently, in vivo analysis of transcriptional termination for the human β-globin gene revealed a new phenomenon—co-transcriptional cleavage (CoTC)3. This primary cleavage event within β-globin pre-messenger RNA, downstream of the poly(A) site, is critical for efficient transcriptional termination by RNA polymerase II3. Here we show that the CoTC process in the human β-globin gene involves an RNA self-cleaving activity. We characterize the autocatalytic core of the CoTC ribozyme and show its functional role in efficient termination in vivo. The identified core CoTC is highly conserved in the 3′ flanking regions of other primate β-globin genes. Functionally, it resembles the 3′ processive, self-cleaving ribozymes described for the protein-encoding genes from the myxomycetes Didymium iridis and Physarum polycephalum, indicating evolutionary conservation of this molecular process. We predict that regulated autocatalytic cleavage elements within pre-mRNAs may be a general phenomenon and that functionally it may provide the entry point for exonucleases involved in mRNA maturation, turnover and, in particular, transcriptional termination.

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Figure 1: Effect of the CoTC element on reconstituted transcription in vitro.
Figure 2: Mapping of the minimal catalytic core within the CoTC element.
Figure 3: Analysis of the CoTC core secondary structure and mapping of the autocatalytic cleavage site.
Figure 4: The catalytic ribozyme core of CoTC mediates termination in vivo and is conserved through evolution.

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Acknowledgements

We thank T. Nilsen, J. Manley, S. Valadkhan, C. Smith and P. C. Branco for their critical comments and constructive support. This work was supported by grants to W.J. from BBSRC and Edward P. Abraham Research Fund, to N.J.P. from the Wellcome Trust and to A.A. from the Wellcome Trust Career Development Programme, Medical Research Council, Cancer Research UK, Edward P. Abraham Research Fund and Exeter College (Oxford).Authors' contributions A.T. is the lead author. A.T.-A. and S.W. are second authors, and contributed equally to this work. A.T.-A. and W.J. are responsible for the RNA secondary structure analysis. S.W. and N.J.P. are responsible for making the CoTC mutants and testing them by nuclear run-on analysis.

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Authors and Affiliations

  1. Sir William Dunn School of Pathology, University of Oxford, South Parks Road, OX1 3RE, Oxford, UK

    Alexandre Teixeira, Abdessamad Tahiri-Alaoui, Steve West, Benjamin Thomas, Igor Martianov, Mick Dye, William James, Nick J. Proudfoot & Alexandre Akoulitchev

  2. The Wellcome Trust Sanger Institute, CB10 1SA, Cambridge, UK

    Aroul Ramadass

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Correspondence to Alexandre Akoulitchev.

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Supplementary information

Supplementary Figure Legends

Legends for Supplementary Figures 1–4. Stability of CoTC RNA. Analysis of the CoTC core secondary structure. Autocatalytic cleavage of the CoTC core and its GTP dependence. (DOC 26 kb)

Supplementary Figures

Supplementary Figures 1–4. Stability of CoTC RNA. Analysis of the CoTC core secondary structure. Autocatalytic cleavage of the CoTC core and its GTP dependence. (PDF 1840 kb)

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Teixeira, A., Tahiri-Alaoui, A., West, S. et al. Autocatalytic RNA cleavage in the human β-globin pre-mRNA promotes transcription termination. Nature 432, 526–530 (2004). https://doi.org/10.1038/nature03032

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