Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Listeria monocytogenes ActA-mediated escape from autophagic recognition

Nature Cell Biology volume 11, pages 1233–1240 (2009)Cite this article

Abstract

Autophagy degrades unnecessary organelles and misfolded protein aggregates1, as well as cytoplasm-invading bacteria2. Nevertheless, the bacteria Listeria monocytogenes efficiently escapes autophagy3,4. We show here that recruitment of the Arp2/3 complex and Ena/VASP, via the bacterial ActA protein, to the bacterial surface disguises the bacteria from autophagic recognition, an activity that is independent of the ability to mediate bacterial motility. L. monocytogenes expressing ActA mutants that lack the ability to recruit the host proteins initially underwent ubiquitylation, followed by recruitment of p62 (also known as SQSTM1) and LC3, before finally undergoing autophagy. The ability of ActA to mediate protection from ubiquitylation was further demonstrated by generating aggregate-prone GFP–ActA–Q79C and GFP–ActA–170* chimaeras, consisting of GFP (green fluorescent protein), the ActA protein and segments of polyQ5 or Golgi membrane protein GCP170 (ref. 6). GFP–ActA–Q79C and GFP–ActA–170* formed aggregates in the host cell cytoplasm, however, these ActA-containing aggregates were not targeted for association with ubiquitin and p62. Our findings indicate that ActA-mediated host protein recruitment is a unique bacterial disguise tactic to escape from autophagy.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: L. monocytogenes lacking ActA is a target of autophagy.
Figure 2: The ability of ActA to mediate bacterial intracellular motility is independent from the ability of L. monocytogenes to escape autophagy.
Figure 3: p62 facilitates autophagy of ubiquitylated ΔactA2.
Figure 4: Anti-aggregating properties of ActA for GFP–polyQ.
Figure 5: Anti-aggregation properties of ActA for GFP–170*.

Similar content being viewed by others

References

  1. Williams, A. et al. Aggregate-prone proteins are cleared from the cytosol by autophagy: therapeutic implications. Curr. Top. Dev. Biol. 76, 89–101 (2006).

    Article  CAS  Google Scholar 

  2. Deretic, V. & Levine, B. Autophagy, immunity, and microbial adaptations. Cell Host Microbe. 5, 527–549 (2009).

    Article  CAS  Google Scholar 

  3. Py, B. F., Lipinski, M. M. & Yuan, J. Autophagy limits Listeria monocytogenes intracellular growth in the early phase of primary infection. Autophagy 3, 117–125 (2007).

    Article  CAS  Google Scholar 

  4. Birmingham, C. L. et al. Listeria monocytogenes evades killing by autophagy during colonization of host cells. Autophagy 3, 442–451 (2007).

    Article  CAS  Google Scholar 

  5. Ikeda, H. et al. Expanded polyglutamine in the Machado-Joseph disease protein induces cell death in vitro and in vivo. Nature Genet. 13, 196–202 (1996).

    Article  CAS  Google Scholar 

  6. Fu, L. et al. Nuclear aggresomes form by fusion of PML-associated aggregates. Mol. Biol. Cell 16, 4905–4917 (2005).

    Article  CAS  Google Scholar 

  7. Klionsky, D. J. & Emr, S. D. Autophagy as a regulated pathway of cellular degradation. Science 290, 1717–1721 (2000).

    Article  CAS  Google Scholar 

  8. Nakagawa, I. et al. Autophagy defends cells against invading group A Streptococcus. Science 306, 1037–1040 (2004).

    Article  CAS  Google Scholar 

  9. Gutierrez, M. G. et al. Autophagy is a defense mechanism inhibiting BCG and Mycobacterium tuberculosis survival in infected macrophages. Cell 119, 753–766 (2004).

    Article  CAS  Google Scholar 

  10. Deretic, V. et al. Mycobacterium tuberculosis inhibition of phagolysosome biogenesis and autophagy as a host defence mechanism. Cell. Microbiol. 8, 719–727 (2006).

    CAS  Google Scholar 

  11. Otto, G. P. et al. Macroautophagy is dispensable for intracellular replication of Legionella pneumophila in Dictyostelium discoideum. Mol. Microbiol. 51, 63–72 (2004).

    CAS  Google Scholar 

  12. Gutierrez, M. G. et al. Autophagy induction favours the generation and maturation of the Coxiella-replicative vacuoles. Cell. Microbiol. 7, 981–993 (2005).

    Article  CAS  Google Scholar 

  13. Dorn, B. R., Dunn, W. A., Jr. & Progulske-Fox, A. Porphyromonas gingivalis traffics to autophagosomes in human coronary artery endothelial cells. Infect. Immun. 69, 5698–5708 (2001).

    Article  CAS  Google Scholar 

  14. Ogawa, M. et al. Escape of intracellular Shigella from autophagy. Science 307, 727–731 (2005).

    Article  CAS  Google Scholar 

  15. Kocks, C. et al. L. monocytogenes-induced actin assembly requires the actA gene product, a surface protein. Cell 68, 521–531 (1992).

    Article  CAS  Google Scholar 

  16. Stevens, J. M., Galyov, E. E. & Stevens, M. P. Actin-dependent movement of bacterial pathogens. Nature Rev. Microbiol. 4, 91–101 (2006).

    Article  CAS  Google Scholar 

  17. Rich, K. A., Burkett, C. & Webster, P. Cytoplasmic bacteria can be targets for autophagy. Cell. Microbiol. 5, 455–468 (2003).

    Article  CAS  Google Scholar 

  18. Birmingham, C. L. et al. Listeriolysin O allows Listeria monocytogenes replication in macrophage vacuoles. Nature 451, 350–354 (2008).

    Article  CAS  Google Scholar 

  19. Welch, M. D., Iwamatsu, A. & Mitchison, T. J. Actin polymerization is induced by Arp2/3 protein complex at the surface of Listeria monocytogenes. Nature 385, 265–269 (1997).

    Article  CAS  Google Scholar 

  20. Chakraborty, T. et al. A focal adhesion factor directly linking intracellularly motile Listeria monocytogenes and Listeria ivanovii to the actin-based cytoskeleton of mammalian cells. EMBO J. 14, 1314–1321 (1995).

    Article  CAS  Google Scholar 

  21. Footer, M. J., Lyo, J. K. & Theriot, J. A. Close packing of Listeria monocytogenes ActA, a natively unfolded protein, enhances F-actin assembly without dimerization. J. Biol. Chem. 283, 23852–23862 (2008).

    Article  CAS  Google Scholar 

  22. Portnoy, D. A., Chakraborty, T., Goebel, W. & Cossart, P. Molecular determinants of Listeria monocytogenes pathogenesis. Infect. Immun. 60, 1263–1267 (1992).

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Pistor, S. et al. Mutations of arginine residues within the 146-KKRRK-150 motif of the ActA protein of Listeria monocytogenes abolish intracellular motility by interfering with the recruitment of the Arp2/3 complex. J. Cell Sci. 113 (Pt 18), 3277–3287 (2000).

    CAS  PubMed  Google Scholar 

  24. Kuma, A. et al. The role of autophagy during the early neonatal starvation period. Nature 432, 1032–1036 (2004).

    Article  CAS  Google Scholar 

  25. Bjorkoy, G. et al. p62/SQSTM1 forms protein aggregates degraded by autophagy and has a protective effect on huntingtin-induced cell death. J. Cell Biol. 171, 603–614 (2005).

    Article  Google Scholar 

  26. Komatsu, M. et al. Homeostatic levels of p62 control cytoplasmic inclusion body formation in autophagy-deficient mice. Cell 131, 1149–1163 (2007).

    Article  CAS  Google Scholar 

  27. Gal, J., Strom, A. L., Kilty, R., Zhang, F. & Zhu, H. p62 accumulates and enhances aggregate formation in model systems of familial amyotrophic lateral sclerosis. J. Biol. Chem. 282, 11068–11077 (2007).

    Article  CAS  Google Scholar 

  28. Pankiv, S. et al. p62/SQSTM1 binds directly to Atg8/LC3 to facilitate degradation of ubiquitylated protein aggregates by autophagy. J. Biol. Chem. 282, 24131–24145 (2007).

    Article  CAS  Google Scholar 

  29. Long, J. et al. Ubiquitin recognition by the UBA domain of p62 involves a novel conformational switch. J. Biol. Chem. 283, 5427–5440 (2008).

    Article  CAS  Google Scholar 

  30. Van Troys, M. et al. The actin propulsive machinery: the proteome of Listeria monocytogenes tails. Biochem. Biophys. Res. Commun. 375, 194–199 (2008).

    Article  CAS  Google Scholar 

  31. Yano, T. et al. Autophagic control of listeria through intracellular innate immune recognition in drosophila. Nature Immunol. 9, 908–916 (2008).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank all members of the Sasakawa laboratory for discussions and technical advice and A. Amend and N. Schlarenko for help with the generation of several ActA mutants. This work was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan to M.O. and C.S, the Special Coordination Funds for Promoting Science from the Japan Science and Technology Agency to C.S, and the ERA-NET Pathogenomics Network funded by the German Ministry of Education and Research and the EU to T.H. and T.C.

Author information

Authors and Affiliations

  1. Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-Ku, 108-8639, Tokyo, Japan

    Yuko Yoshikawa, Michinaga Ogawa, Makoto Fukumatsu, Hitomi Mimuro & Chihiro Sasakawa

  2. Institute of Medical Microbiology, Justus-Liebig University Giessen, Frankfurter Strasse 107, Giessen, D-35392, Germany

    Torsten Hain & Trinad Chakraborty

  3. Division of Ultrastructual Research, BioMedical Research Center, Graduate School of Medicine, Juntendo University, 2-1-1, Hongo, Bunkyo-ku, 113-8421, Tokyo, Japan

    Mitsutaka Yoshida

  4. Department of Infectious Disease Control, International Research Center for Infectious Diseases, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-Ku, 108-8639, Tokyo, Japan

    Minsoo Kim, Ichiro Nakagawa & Chihiro Sasakawa

  5. Graduate School of Comprehensive Human Sciences, University of Tsukuba, 1-1-1, Tennodai, Tsukuba, 305-8575, Ibaraki, Japan

    Toru Yanagawa & Tetsuro Ishii

  6. Kyoto University Graduate School of Biostudies and Solution Oriented Research for Science and Technology (JST), Kyoto, 606-8501, Japan

    Akira Kakizuka

  7. Department of Cell Biology, University of Alabama at Birmingham, Birmingham, 35294, AL, USA

    Elizabeth Sztul

  8. CREST, Japan Science and Technology Agency, Kawaguchi, 332–0012, Japan

    Chihiro Sasakawa

Authors
  1. Yuko Yoshikawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Michinaga Ogawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Torsten Hain
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mitsutaka Yoshida
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Makoto Fukumatsu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Minsoo Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Hitomi Mimuro
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Ichiro Nakagawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Toru Yanagawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Tetsuro Ishii
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Akira Kakizuka
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Elizabeth Sztul
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Trinad Chakraborty
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Chihiro Sasakawa
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

C.S. and T.C. conceived the study. Y.Y. and M.O. performed experiments. Y.Y., M.O., C.S. and T.C. analysed data. M.F., K.M., M.H. and I.N. provided technical advice. M.Y. performed electon microscopic analysis. T.Y. and T.I. generated p62-null mouse embryonic fibroblasts. A.K. generated the Q79C plasmid. E.S. generated the GFP-170* plasmid. T.H. and T.C. generated bacterial strains and antibodies. C.S. and T.C. wrote the manuscript.

Corresponding author

Correspondence to Chihiro Sasakawa.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

Supplementary Information (PDF 3672 kb)

Supplementary Information

Supplementary Movie 1 (MOV 4018 kb)

Supplementary Information

Supplementary Movie 2 (MOV 8135 kb)

Supplementary Information

Supplementary Movie 3 (MOV 3769 kb)

Supplementary Information

Supplementary Movie 4 (MOV 8404 kb)

Supplementary Information

Supplementary Movie 5 (MOV 3466 kb)

Supplementary Information

Supplementary Movie 6 (MOV 10188 kb)

Supplementary Information

Supplementary Movie 7 (MOV 3634 kb)

Supplementary Information

Supplementary Movie 8 (MOV 7396 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Yoshikawa, Y., Ogawa, M., Hain, T. et al. Listeria monocytogenes ActA-mediated escape from autophagic recognition. Nat Cell Biol 11, 1233–1240 (2009). https://doi.org/10.1038/ncb1967

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ncb1967

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing