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Legionella pneumophila SidD is a deAMPylase that modifies Rab1

Nature volume 475, pages 506–509 (2011)Cite this article

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Abstract

Legionella pneumophila actively modulates host vesicle trafficking pathways to facilitate its intracellular replication with effectors translocated by the Dot/Icm type IV secretion system (T4SS)1. The SidM/DrrA protein functions by locking the small GTPase Rab1 into an active form by its guanine nucleotide exchange factor (GEF) and AMPylation activity2,3,4. Here we demonstrate that the L. pneumophila protein SidD preferably deAMPylates Rab1. We found that the deAMPylation activity of SidD could suppress the toxicity of SidM to yeast and is required to release Rab1 from bacterial phagosomes efficiently. A molecular mechanism for the temporal control of Rab1 activity in different phases of L. pneumophila infection is thus established. These observations indicate that AMPylation-mediated signal transduction is a reversible process regulated by specific enzymes.

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Figure 1: Suppression of the cytotoxicity of SidM by SidD.
Figure 2: SidD is a deAMPylase that targets SidM-modified Rab1.
Figure 3: The Asp residue at position 92 or 110 is important for SidD activity.
Figure 4: SidD is required for efficient removal of Rab1 from L. pneumophila phagosome.

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References

  1. Ensminger, A. W. & Isberg, R. R. Legionella pneumophila Dot/Icm translocated substrates: a sum of parts. Curr. Opin. Microbiol. 12, 67–73 (2009)

    Article  CAS  Google Scholar 

  2. Machner, M. P. & Isberg, R. R. Targeting of host Rab GTPase function by the intravacuolar pathogen Legionella pneumophila . Dev. Cell 11, 47–56 (2006)

    Article  CAS  Google Scholar 

  3. Murata, T. et al. The Legionella pneumophila effector protein DrrA is a Rab1 guanine nucleotide-exchange factor. Nature Cell Biol. 8, 971–977 (2006)

    Article  CAS  Google Scholar 

  4. Muller, M. P. et al. The Legionella effector protein DrrA AMPylates the membrane traffic regulator Rab1b. Science 329, 946–949 (2010)

    Article  ADS  Google Scholar 

  5. Isberg, R. R., O'Connor, T. J. & Heidtman, M. The Legionella pneumophila replication vacuole: making a cosy niche inside host cells. Nature Rev. Microbiol. 7, 13–24 (2009)

    Article  CAS  Google Scholar 

  6. Kubori, T., Shinzawa, N., Kanuka, H. & Nagai, H. Legionella metaeffector exploits host proteasome to temporally regulate cognate effector. PLoS Pathog. 6, e1001216 (2010)

    Article  Google Scholar 

  7. Ingmundson, A., Delprato, A., Lambright, D. G. & Roy, C. R. Legionella pneumophila proteins that regulate Rab1 membrane cycling. Nature 450, 365–369 (2007)

    Article  ADS  CAS  Google Scholar 

  8. Worby, C. A. et al. The fic domain: regulation of cell signaling by adenylylation. Mol. Cell 34, 93–103 (2009)

    Article  CAS  Google Scholar 

  9. Kinch, L. N., Yarbrough, M. L., Orth, K. & Grishin, N. V. Fido, a novel AMPylation domain common to Fic, Doc, and AvrB. PLoS ONE 4, e5818 (2009)

    Article  ADS  Google Scholar 

  10. Pan, X., Luhrmann, A., Satoh, A., Laskowski-Arce, M. A. & Roy, C. R. Ankyrin repeat proteins comprise a diverse family of bacterial type IV effectors. Science 320, 1651–1654 (2008)

    Article  ADS  CAS  Google Scholar 

  11. Belyi, Y., Tabakova, I., Stahl, M. & Aktories, K. Lgt: a family of cytotoxic glucosyltransferases produced by Legionella pneumophila . J. Bacteriol. 190, 3026–3035 (2008)

    Article  CAS  Google Scholar 

  12. Shen, X. et al. Targeting eEF1A by a Legionella pneumophila effector leads to inhibition of protein synthesis and induction of host stress response. Cell. Microbiol. 11, 911–926 (2009)

    Article  CAS  Google Scholar 

  13. Luo, Z. Q. & Isberg, R. R. Multiple substrates of the Legionella pneumophila Dot/Icm system identified by interbacterial protein transfer. Proc. Natl Acad. Sci. USA 101, 841–846 (2004)

    Article  ADS  CAS  Google Scholar 

  14. Chien, M. et al. The genomic sequence of the accidental pathogen Legionella pneumophila . Science 305, 1966–1968 (2004)

    Article  ADS  CAS  Google Scholar 

  15. Roy, C. R. & Mukherjee, S. Bacterial FIC proteins AMP up infection. Sci. Signal. 2, pe14 (2009)

    Article  Google Scholar 

  16. Brombacher, E. et al. Rab1 guanine nucleotide exchange factor SidM is a major phosphatidylinositol 4-phosphate-binding effector protein of Legionella pneumophila . J. Biol. Chem. 284, 4846–4856 (2009)

    Article  CAS  Google Scholar 

  17. Soding, J. Protein homology detection by HMM–HMM comparison. Bioinformatics 21, 951–960 (2005)

    Article  Google Scholar 

  18. Rantanen, M. K., Lehtio, L., Rajagopal, L., Rubens, C. E. & Goldman, A. Structure of Streptococcus agalactiae serine/threonine phosphatase. The subdomain conformation is coupled to the binding of a third metal ion. FEBS J. 274, 3128–3137 (2007)

    Article  CAS  Google Scholar 

  19. Schlicker, C. et al. Structural analysis of the PP2C phosphatase tPphA from Thermosynechococcus elongatus: a flexible flap subdomain controls access to the catalytic site. J. Mol. Biol. 376, 570–581 (2008)

    Article  CAS  Google Scholar 

  20. Seabra, M. C. & Wasmeier, C. Controlling the location and activation of Rab GTPases. Curr. Opin. Cell Biol. 16, 451–457 (2004)

    Article  CAS  Google Scholar 

  21. Woolery, A. R., Luong, P., Broberg, C. A. & Orth, K. AMPylation: something old is new again. Front. Microbiol 1, 113 (2010)

    Article  CAS  Google Scholar 

  22. Yarbrough, M. L. et al. AMPylation of Rho GTPases by Vibrio VopS disrupts effector binding and downstream signaling. Science 323, 269–272 (2009)

    Article  CAS  Google Scholar 

  23. Zhu, W. et al. Comprehensive identification of protein substrates of the Dot/Icm type IV transporter of Legionella pneumophila . PLoS ONE 6, e17638 (2011)

    Article  ADS  CAS  Google Scholar 

  24. Berger, K. H. & Isberg, R. R. Two distinct defects in intracellular growth complemented by a single genetic locus in Legionella pneumophila . Mol. Microbiol. 7, 7–19 (1993)

    Article  CAS  Google Scholar 

  25. Conover, G. M., Derre, I., Vogel, J. P. & Isberg, R. R. The Legionella pneumophila LidA protein: a translocated substrate of the Dot/Icm system associated with maintenance of bacterial integrity. Mol. Microbiol. 48, 305–321 (2003)

    Article  CAS  Google Scholar 

  26. Liu, Y., Gao, P., Banga, S. & Luo, Z. Q. An in vivo gene deletion system for determining temporal requirement of bacterial virulence factors. Proc. Natl Acad. Sci. USA 105, 9385–9390 (2008)

    Article  ADS  CAS  Google Scholar 

  27. Fan, H. Y., Cheng, K. K. & Klein, H. L. Mutations in the RNA polymerase II transcription machinery suppress the hyperrecombination mutant hpr1Δ of Saccharomyces cerevisiae . Genetics 142, 749–759 (1996)

    CAS  PubMed  PubMed Central  Google Scholar 

  28. Gietz, R. D., Schiestl, R. H., Willems, A. R. & Woods, R. A. Studies on the transformation of intact yeast cells by the LiAc/SS-DNA/PEG procedure. Yeast 11, 355–360 (1995)

    Article  CAS  Google Scholar 

  29. Xu, L. et al. Inhibition of host vacuolar H+-ATPase activity by a Legionella pneumophila effector. PLoS Pathog. 6, e1000822 (2010)

    Article  Google Scholar 

Download references

Acknowledgements

We thank R. Isberg for the antibody against SidM and A. Aronson and A. Tao for critical reading of the manuscript and for discussions. This work was supported by NIH-NIAID grants R01AI069344, K02AI085403 and R21AI092043 (Z.-Q.L).

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  1. Department of Biological Sciences, Purdue University, 915 West State Street, West Lafayette, Indiana 47907, USA,

    Yunhao Tan & Zhao-Qing Luo

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  1. Yunhao Tan
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Contributions

Y.T. and Z.-Q.L. conceived the project. Y.T. performed the experiments. Y.T. and Z.-Q.L analysed the data. Z.-Q.L wrote the paper.

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Correspondence to Zhao-Qing Luo.

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The authors declare no competing financial interests.

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Tan, Y., Luo, ZQ. Legionella pneumophila SidD is a deAMPylase that modifies Rab1. Nature 475, 506–509 (2011). https://doi.org/10.1038/nature10307

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