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RNA-mediated interaction between the peptide-binding and GTPase domains of the signal recognition particle

Nature Structural & Molecular Biology volume 12, pages 11161122 (2005) | Download Citation

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Abstract

The signal recognition particle (SRP) targets nascent proteins to cellular membranes for insertion or secretion by recognizing polypeptides containing an N-terminal signal sequence as they emerge from the ribosome. GTP-dependent binding of SRP to its receptor protein leads to controlled release of the nascent chain into a membrane-spanning translocon pore. Here we show that the association of the SRP with its receptor triggers a marked conformational change in the complex, localizing the SRP RNA and the adjacent signal peptide–binding site at the SRP-receptor heterodimer interface. The orientation of the RNA suggests how peptide binding and GTP hydrolysis can be coupled through direct structural contact during cycles of SRP-directed protein translocation.

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Acknowledgements

We thank I.J. MacRae for helpful discussions on the BABE-Fe modification sites of Ffh and FtsY and K. Karbstein, W. Gilbert, C.S. Fraser, N.H. Chmiel, J.W. Hershey and H.F. Noller for review of the manuscript. The work was supported by grant GM22778 from the US National Institutes of Health.

Author information

Affiliations

  1. Department of Molecular and Cell Biology, University of California, Berkeley, California 94705, USA.

    • Richard J Spanggord
    • , Fai Siu
    • , Ailong Ke
    •  & Jennifer A Doudna
  2. Department of Chemistry, University of California, Berkeley, California 94705, USA.

    • Jennifer A Doudna
  3. Howard Hughes Medical Institute, University of California, Berkeley, California 94705, USA.

    • Jennifer A Doudna
  4. Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA.

    • Jennifer A Doudna

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Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Jennifer A Doudna.

Supplementary information

PDF files

  1. 1.

    Supplementary Fig. 1

    Fluorescence assay to monitor BABE-Fe conjugation of Ffh and FtsY.

  2. 2.

    Supplementary Fig. 2

    Gel mobility shift assay of BABE-Fe–modified Ffh and FtsY.

  3. 3.

    Supplementary Fig. 3

    Ffh M- and G-domain cleavage quantification.

  4. 4.

    Supplementary Fig. 4

    View of the Ffh-FtsY heterodime.r

  5. 5.

    Supplementary Fig. 5

    FtsY cleavage data quantification.

  6. 6.

    Supplementary Fig. 6

    Molecular models of the SRP-FtsY complex.

  7. 7.

    Supplementary Fig. 7

    Quantification of cleavage data produced by Ffh.

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DOI

https://doi.org/10.1038/nsmb1025

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