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Structural analysis of cooperative RNA binding by the La motif and central RRM domain of human La protein

Nature Structural & Molecular Biology volume 11, pages 323–329 (2004)Cite this article

Abstract

The La protein is a conserved component of eukaryotic ribonucleoprotein complexes that binds the 3′ poly(U)-rich elements of nascent RNA polymerase III (pol III) transcripts to assist folding and maturation. This specific recognition is mediated by the N-terminal domain (NTD) of La, which comprises a La motif and an RNA recognition motif (RRM). We have determined the solution structures of both domains and show that the La motif adopts an α/β fold that comprises a winged-helix motif elaborated by the insertion of three helices. Chemical shift mapping experiments show that these insertions are involved in RNA interactions. They further delineate a distinct surface patch on each domain—containing both basic and aromatic residues—that interacts with RNA and accounts for the cooperative binding of short oligonucleotides exhibited by the La NTD.

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Figure 1: Domain organization of human La and deletion mutants used in this study.
Figure 2: Structural analysis of the La motif and the adjacent RRM.
Figure 3: Analysis of La-oligo(U) interaction.
Figure 4: Alignment of the La motif and central RRM sequences for La homologs.

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Acknowledgements

We are grateful to D. Kenan, J. Keene, N. Sonenberg and R. Maraia for the gift of plasmids containing constructs of human La. We thank S. Wolin and K. Reinisch for communicating results before publication. We are indebted to The Wellcome Trust for financial support. C.A. and D.S. are Institute of Biomedical and Biomolecular Sciences research students. A list of NMR assignments and restraints is available from M.R.C. (sasi.conte@port.ac.uk).

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Author notes

  1. Amanda Jacks

    Present address: Department of Biochemistry and Biophysics, Texas A&M University, MS 2128, College Station, Texas, 77843-2128, USA

  2. Caterina Alfano and Domenico Sanfelice: These authors contributed equally to this work.

Authors and Affiliations

  1. Biophysics Laboratories, Institute of Biomedical and Biomolecular Sciences, University of Portsmouth, St. Michael's Building, Portsmouth, PO1 2DT, UK

    Caterina Alfano, Domenico Sanfelice, Amanda Jacks & Maria R Conte

  2. Department of Molecular Structure, Walter and Eliza Hall Institute, Parkville, 3052, Victoria, Australia

    Jeff Babon

  3. Biomedical NMR Centre, National Institute for Medical Research, The Ridgeway, Mill Hill, London, NW7 1AA, UK

    Geoff Kelly

  4. Department of Biological Sciences, Biophysics Section, Blackett Laboratory, Imperial College London, London, SW7 2AZ, UK

    Stephen Curry

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Correspondence to Maria R Conte.

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Alfano, C., Sanfelice, D., Babon, J. et al. Structural analysis of cooperative RNA binding by the La motif and central RRM domain of human La protein. Nat Struct Mol Biol 11, 323–329 (2004). https://doi.org/10.1038/nsmb747

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