Cryo-EM structure of the E. coli translating ribosome in complex with SRP and its receptor

Article metrics

Abstract

We report the 'early' conformation of the Escherichia coli signal recognition particle (SRP) and its receptor FtsY bound to the translating ribosome, as determined by cryo-EM. FtsY binds to the tetraloop of the SRP RNA, whereas the NG domains of the SRP protein and FtsY interact weakly in this conformation. Our results suggest that optimal positioning of the SRP RNA tetraloop and the Ffh NG domain leads to FtsY recruitment.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: Generation and characterization of RNC–SRP–FtsY.
Figure 2: Atomic model of the early conformation of scSRP.
Figure 3: Cartoon model of co-translational targeting.

Accession codes

Primary accessions

Electron Microscopy Data Bank

Protein Data Bank

References

  1. 1

    Walter, P. & Johnson, A.E. Annu. Rev. Cell Biol. 10, 87–119 (1994).

  2. 2

    Nagai, K. et al. EMBO J. 22, 3479–3485 (2003).

  3. 3

    Doudna, J.A. & Batey, R.T. Annu. Rev. Biochem. 73, 539–557 (2004).

  4. 4

    Ulbrandt, N.D., Newitt, J.A. & Bernstein, H.D. Cell 88, 187–196 (1997).

  5. 5

    Freymann, D.M., Keenan, R.J., Stroud, R.M. & Walter, P. Nature 385, 361–364 (1997).

  6. 6

    Montoya, G., Svensson, C., Luirink, J. & Sinning, I. Nature 385, 365–368 (1997).

  7. 7

    Connolly, T., Rapiejko, P. & Gilmore, R. Science 252, 1171–1173 (1991).

  8. 8

    Focia, P.J., Shepotinovskaya, I.V., Seidler, J.A. & Freymann, D.M. Science 303, 373–377 (2004).

  9. 9

    Egea, P.F. et al. Nature 427, 215–221 (2004).

  10. 10

    Batey, R.T. et al. Science 287, 1232–1239 (2000).

  11. 11

    Weiche, B. et al. J. Mol. Biol. 377, 761–773 (2008).

  12. 12

    Zhang, X., Schaffitzel, C., Ban, N. & Shan, S.O. Proc. Natl. Acad. Sci. USA 106, 1754–1759 (2009).

  13. 13

    Schaffitzel, C. et al. Nature 444, 503–506 (2006).

  14. 14

    Halic, M. et al. Nature 444, 507–511 (2006).

  15. 15

    Schuwirth, B.S. et al. Science 310, 827–834 (2005).

  16. 16

    Shen, K. & Shan, S.O. Proc. Natl. Acad. Sci. USA 107, 7698–7703 (2010).

  17. 17

    Jagath, J.R. et al. RNA 7, 293–301 (2001).

  18. 18

    Siu, F.Y., Spanggord, R.J. & Doudna, J.A. RNA 13, 240–250 (2007).

  19. 19

    Halic, M. et al. Science 312, 745–747 (2006).

  20. 20

    Bradshaw, N., Neher, S.B., Booth, D.S. & Walter, P. Science 323, 127–130 (2009).

Download references

Acknowledgements

We would like to thank G. Schoehn for collecting EM data, I. Berger for designing scSRP, C. Frick for technical assistance and X. Zhang for helpful suggestions. W. Wintermeyer (Max Planck Institute for Biophysical Chemistry) kindly provided pET24a-Ffh. J. Luirink (Vrije Universiteit Amsterdam) is acknowledged for providing pET9-FtsY. This work was supported by the Swiss National Science Foundation (SNSF) and the National Center of Excellence in Research Structural Biology program of the SNSF. The authors acknowledge support by the Electron Microscopy of ETH Zurich (EMEZ) and from the infrastructure of the Partnership for Structural Biology in Grenoble.

Author information

C.S. conceived the scSRP construct and performed sample preparations; S.-o.S. performed activity assays; D.B. did the electron microscopy; D.B., L.F.E. and C.S. performed the image analysis and model building; C.S., N.B., D.B. and S.-o.S. prepared the manuscript.

Correspondence to Nenad Ban or Christiane Schaffitzel.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–5, Supplementary Methods and Supplementary Discussion (PDF 738 kb)

Rights and permissions

Reprints and Permissions

About this article

Further reading