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Structure-function analysis of hRPC62 provides insights into RNA polymerase III transcription initiation

Nature Structural & Molecular Biology volume 18, pages 352–358 (2011)Cite this article

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A Corrigendum to this article was published on 06 April 2011

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Abstract

The 17-subunit human RNA polymerase III (hPol III) transcribes small, untranslated RNA genes that are involved in the regulation of transcription, splicing and translation. hPol III subunits hRPC62, hRPC39 and hRPC32 form a stable ternary subcomplex required for promoter-specific transcription initiation by hPol III. Here, we report the crystal structure of hRPC62. This subunit folds as a four-tandem extended winged helix (eWH) protein that is structurally related to the transcription factor TFIIEα N terminus. Through biochemical analyses, we mapped the protein-protein interactions of hRPC62, hRPC32 and hRPC39. In addition, we demonstrated that hRPC62 and hRPC39 bind single-stranded and duplex DNA, respectively, in a sequence-independent manner. Overall, we shed light on structural similarities between the hPol III–specific subunit hRPC62 and TFIIEα and propose specific functions for hRPC39 and hRPC62 in transcription initiation by hPol III.

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Figure 1: Overall structure of hRPC62.
Figure 2: Interaction mapping of the two isoforms of hRPC32 with hRPC62.
Figure 3: Mutual-interaction mapping of hRPC39 and hRPC62.
Figure 4: DNA-binding properties of hRPC62.
Figure 5: DNA-binding properties of hRPC39.
Figure 6: hRPC62 (Rpc82) positioning into the yeast RNA Pol III electron-density map.

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  • 17 March 2011

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Acknowledgements

We acknowledge the European Synchrotron Radiation Facility for provision of synchrotron radiation facilities, and we would like to thank staff members for assistance in using beamline ID14-1 and ID29. We also acknowledge the Synchrotron SOLEIL for providing access to Proxima-1 beamline. We are grateful to M. Werner and A. Delprato for critical reading of the manuscript and C. Monfoulet for technical assistance. This work was supported by an internal grant from the Institut Européen de Chimie et Biologie and the University of Bordeaux 2 (S.F. and M.T.) and by grants from the Ligue National Contre le Cancer, comités de Gironde et Dordogne (to M.T.). S.L. was supported by a PhD fellowship from the Conseil Régional d'Aquitaine. The authors declare that they have no competing financial interests.

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Authors and Affiliations

  1. Institut Européen de Chimie et Biologie, Pessac, France

    Stéphane Lefèvre, Hélène Dumay-Odelot, Leyla El-Ayoubi, Noël Pinaud, Martin Teichmann & Sébastien Fribourg

  2. Inserm and Université de Bordeaux, Bordeaux, France

    Stéphane Lefèvre, Hélène Dumay-Odelot, Leyla El-Ayoubi, Noël Pinaud, Martin Teichmann & Sébastien Fribourg

  3. European Molecular Biology Laboratory, Heidelberg, Germany

    Aidan Budd

  4. Synchrotron SOLEIL, St-Aubin, France

    Pierre Legrand

  5. CESAMO ISM, Université de Bordeaux, Pessac, France

    Noël Pinaud

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Contributions

S.L. and N.P. purified and crystallized hRPC62. S.L., P.L. and S.F. determined the crystal structure of hRPC62. A.B. performed the bioinformatics analysis. H.D.-O., L.E.-A. and S.F. conducted functional assays. M.T. contributed to manuscript preparation. S.F. designed, supervised research and wrote the manuscript.

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Correspondence to Sébastien Fribourg.

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Lefèvre, S., Dumay-Odelot, H., El-Ayoubi, L. et al. Structure-function analysis of hRPC62 provides insights into RNA polymerase III transcription initiation. Nat Struct Mol Biol 18, 352–358 (2011). https://doi.org/10.1038/nsmb.1996

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