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RNA polymerase II pauses and associates with pre-mRNA processing factors at both ends of genes

Nature Structural & Molecular Biology volume 15, pages 7178 (2008) | Download Citation

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Abstract

We investigated co-transcriptional recruitment of pre-mRNA processing factors to human genes. Capping factors associate with paused RNA polymerase II (pol II) at the 5′ ends of quiescent genes. They also track throughout actively transcribed genes and accumulate with paused polymerase in the 3′ flanking region. The 3′ processing factors cleavage stimulation factor and cleavage polyadenylation specificity factor are maximally recruited 0.5–1.5 kilobases downstream of poly(A) sites where they coincide with capping factors, Spt5, and Ser2-hyperphosphorylated, paused pol II. 3′ end processing factors also localize at transcription start sites, and this early recruitment is enhanced after polymerase arrest with the elongation factor DRB. These results suggest that promoters may help specify recruitment of 3′ end processing factors. We propose a dual-pausing model wherein elongation arrests near the transcription start site and in the 3′ flank to allow co-transcriptional processing by factors recruited to the pol II ternary complex.

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Acknowledgements

This work was supported by US National Institutes of Health (NIH) grants GM063873 and GM58613 to D.L.B. and CA117907 to J.E. K.G.-C. was supported by NIH fellowship 5F31 GM072099 and S.K. by F32 GM076951. We thank I. Mattaj (EMBL, Heidelberg) and T. Blumenthal (University of Colorado, Boulder) for antibodies. We also thank T. Blumenthal, J. Jaehning, R. Davis, T. Evans, N. Gomes, G. Bjerke, G. Bilousova, B. Erickson and members of the Bentley and Espinosa labs for their help.

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Author notes

    • Kira Glover-Cutter
    •  & Soojin Kim

    These authors contributed equally to this work.

Affiliations

  1. Department of Biochemistry and Molecular Genetics, University of Colorado School of Medicine, UCHSC, MS8101, PO Box 6511, Aurora, Colorado 80045, USA.

    • Kira Glover-Cutter
    • , Soojin Kim
    •  & David L Bentley
  2. Department of Molecular Cellular and Developmental Biology, University of Colorado, Boulder, Colorado 80309, USA.

    • Joaquin Espinosa

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Correspondence to David L Bentley.

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DOI

https://doi.org/10.1038/nsmb1352

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