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Listeria monocytogenes ActA-mediated escape from autophagic recognition

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.

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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*.

References

  1. 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).

    CAS  Article  Google Scholar 

  2. 2

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

    CAS  Article  Google Scholar 

  3. 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).

    CAS  Article  Google Scholar 

  4. 4

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

    CAS  Article  Google Scholar 

  5. 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).

    CAS  Article  Google Scholar 

  6. 6

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

    CAS  Article  Google Scholar 

  7. 7

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

    CAS  Article  Google Scholar 

  8. 8

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

    CAS  Article  Google Scholar 

  9. 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).

    CAS  Article  Google Scholar 

  10. 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. 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. 12

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

    CAS  Article  Google Scholar 

  13. 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).

    CAS  Article  Google Scholar 

  14. 14

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

    CAS  Article  Google Scholar 

  15. 15

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

    CAS  Article  Google Scholar 

  16. 16

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

    CAS  Article  Google Scholar 

  17. 17

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

    CAS  Article  Google Scholar 

  18. 18

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

    CAS  Article  Google Scholar 

  19. 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).

    CAS  Article  Google Scholar 

  20. 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).

    CAS  Article  Google Scholar 

  21. 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).

    CAS  Article  Google Scholar 

  22. 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. 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. 24

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

    CAS  Article  Google Scholar 

  25. 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. 26

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

    CAS  Article  Google Scholar 

  27. 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).

    CAS  Article  Google Scholar 

  28. 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).

    CAS  Article  Google Scholar 

  29. 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).

    CAS  Article  Google Scholar 

  30. 30

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

    CAS  Article  Google Scholar 

  31. 31

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

    CAS  Article  Google Scholar 

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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.

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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.

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Correspondence to Chihiro Sasakawa.

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The authors declare no competing financial interests.

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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

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