L. monocytogenes is a facultative intracellular bacterium responsible for listeriosis. It is able to invade, survive and replicate in phagocytic and non-phagocytic cells. The infectious process at the cellular level has been extensively studied and many virulence factors have been identified. Yet, the role of InlK, a member of the internalin family specific to L. monocytogenes, remains unknown. Here, we first show using deletion analysis and in vivo infection, that InlK is a bona fide virulence factor, poorly expressed in vitro and well expressed in vivo, and that it is anchored to the bacterial surface by sortase A. We then demonstrate by a yeast two hybrid screen using InlK as a bait, validated by pulldown experiments and immunofluorescence analysis that intracytosolic bacteria via an interaction with the protein InlK interact with the Major Vault Protein (MVP), the main component of cytoplasmic ribonucleoproteic particules named vaults. Although vaults have been implicated in several cellular processes, their role has remained elusive. Our analysis demonstrates that MVP recruitment disguises intracytosolic bacteria from autophagic recognition, leading to an increased survival rate of InlK over-expressing bacteria compared to InlK? bacteria. Together these results reveal that MVP is hijacked by L. monocytogenes in order to counteract the autophagy process, a finding that could have major implications in deciphering the cellular role of vault particles.
References
[1]
Dortet L, Veiga-Chacon E, Cossart P (2009) Listeria monocytogenes. In: Schaechter M, editor. Encyclopedia of Microbiology. 3rd ed. Oxford: Elsevier. pp. 182–198.
[2]
Disson O, Grayo S, Huillet E, Nikitas G, Langa-Vives F, et al. (2008) Conjugated action of two species-specific invasion proteins for fetoplacental listeriosis. Nature 455: 1114–1118.
[3]
Hamon M, Bierne H, Cossart P (2006) Listeria monocytogenes: a multifaceted model. Nat Rev Microbiol 4: 423–434.
[4]
Lecuit M (2007) Human listeriosis and animal models. Microbes Infect 9: 1216–1225.
[5]
Cossart P, Sansonetti PJ (2004) Bacterial invasion: the paradigms of enteroinvasive pathogens. Science 304: 242–248.
[6]
Bierne H, Gouin E, Roux P, Caroni P, Yin HL, et al. (2001) A role for cofilin and LIM kinase in Listeria-induced phagocytosis. J Cell Biol 155: 101–112.
[7]
Bonazzi M, Veiga E, Pizarro-Cerda J, Cossart P (2008) Successive post-translational modifications of E-cadherin are required for InlA-mediated internalization of Listeria monocytogenes. Cell Microbiol 10: 2208–2222.
[8]
Kocks C, Gouin E, Tabouret M, Berche P, Ohayon H, et al. (1992) L. monocytogenes-induced actin assembly requires the actA gene product, a surface protein. Cell 68: 521–531.
[9]
Mostowy S, Cossart P (2011) Autophagy and the cytoskeleton: New links revealed by intracellular pathogens. Autophagy 7: 780–782.
[10]
Cossart P (2000) Actin-based motility of pathogens: the Arp2/3 complex is a central player. Cell Microbiol 2: 195–205.
[11]
Yoshikawa Y, Ogawa M, Hain T, Yoshida M, Fukumatsu M, et al. (2009) Listeria monocytogenes ActA-mediated escape from autophagic recognition. Nat Cell Biol 11: 1233–1240.
[12]
Thurston TL, Ryzhakov G, Bloor S, von Muhlinen N, Randow F (2009) The TBK1 adaptor and autophagy receptor NDP52 restricts the proliferation of ubiquitin-coated bacteria. Nat Immunol 10: 1215–1221.
[13]
Birmingham CL, Canadien V, Gouin E, Troy EB, Yoshimori T, et al. (2007) Listeria monocytogenes evades killing by autophagy during colonization of host cells. Autophagy 3: 442–451.
[14]
Perrin AJ, Jiang X, Birmingham CL, So NS, Brumell JH (2004) Recognition of bacteria in the cytosol of Mammalian cells by the ubiquitin system. Curr Biol 14: 806–811.
[15]
Bierne H, Cossart P (2007) Listeria monocytogenes surface proteins: from genome predictions to function. Microbiol Mol Biol Rev 71: 377–397.
[16]
Tanaka H, Kato K, Yamashita E, Sumizawa T, Zhou Y, et al. (2009) The structure of rat liver vault at 3.5 angstrom resolution. Science 323: 384–388.
[17]
Rome L, Kedersha N, Chugani D (1991) Unlocking vaults: organelles in search of a function. Trends Cell Biol 1: 47–50.
[18]
Anderson DH, Kickhoefer VA, Sievers SA, Rome LH, Eisenberg D (2007) Draft crystal structure of the vault shell at 9-A resolution. PLoS Biol 5: e318.
[19]
Mikyas Y, Makabi M, Raval-Fernandes S, Harrington L, Kickhoefer VA, et al. (2004) Cryoelectron microscopy imaging of recombinant and tissue derived vaults: localization of the MVP N termini and VPARP. J Mol Biol 344: 91–105.
[20]
Stephen AG, Raval-Fernandes S, Huynh T, Torres M, Kickhoefer VA, et al. (2001) Assembly of vault-like particles in insect cells expressing only the major vault protein. J Biol Chem 276: 23217–23220.
[21]
Kickhoefer VA, Liu Y, Kong LB, Snow BE, Stewart PL, et al. (2001) The Telomerase/vault-associated protein TEP1 is required for vault RNA stability and its association with the vault particle. J Cell Biol 152: 157–164.
[22]
Kickhoefer VA, Siva AC, Kedersha NL, Inman EM, Ruland C, et al. (1999) The 193-kD vault protein, VPARP, is a novel poly(ADP-ribose) polymerase. J Cell Biol 146: 917–928.
[23]
Liu Y, Snow BE, Kickhoefer VA, Erdmann N, Zhou W, et al. (2004) Vault poly(ADP-ribose) polymerase is associated with mammalian telomerase and is dispensable for telomerase function and vault structure in vivo. Mol Cell Biol 24: 5314–5323.
[24]
Raval-Fernandes S, Kickhoefer VA, Kitchen C, Rome LH (2005) Increased susceptibility of vault poly(ADP-ribose) polymerase-deficient mice to carcinogen-induced tumorigenesis. Cancer Res 65: 8846–8852.
van Zon A, Mossink MH, Schoester M, Scheffer GL, Scheper RJ, et al. (2001) Multiple human vault RNAs. Expression and association with the vault complex. J Biol Chem 276: 37715–37721.
[27]
Berger W, Steiner E, Grusch M, Elbling L, Micksche M (2009) Vaults and the major vault protein: novel roles in signal pathway regulation and immunity. Cell Mol Life Sci 66: 43–61.
[28]
Steiner E, Holzmann K, Elbling L, Micksche M, Berger W (2006) Cellular functions of vaults and their involvement in multidrug resistance. Curr Drug Targets 7: 923–934.
[29]
Schroeijers AB, Reurs AW, Scheffer GL, Stam AG, de Jong MC, et al. (2002) Up-regulation of drug resistance-related vaults during dendritic cell development. J Immunol 168: 1572–1578.
[30]
Kowalski MP, Dubouix-Bourandy A, Bajmoczi M, Golan DE, Zaidi T, et al. (2007) Host resistance to lung infection mediated by major vault protein in epithelial cells. Science 317: 130–132.
[31]
Mrazek J, Kreutmayer SB, Grasser FA, Polacek N, Huttenhofer A (2007) Subtractive hybridization identifies novel differentially expressed ncRNA species in EBV-infected human B cells. Nucleic Acids Res 35: e73.
[32]
Kim E, Lee S, Mian MF, Yun SU, Song M, et al. (2006) Crosstalk between Src and major vault protein in epidermal growth factor-dependent cell signalling. FEBS J 273: 793–804.
[33]
Kolli S, Zito CI, Mossink MH, Wiemer EA, Bennett AM (2004) The major vault protein is a novel substrate for the tyrosine phosphatase SHP-2 and scaffold protein in epidermal growth factor signaling. J Biol Chem 279: 29374–29385.
[34]
Liang P, Wan Y, Yan Y, Wang Y, Luo N, et al. (2010) MVP interacts with YPEL4 and inhibits YPEL4-mediated activities of the ERK signal pathway. Biochem Cell Biol 88: 445–450.
[35]
Yu Z, Fotouhi-Ardakani N, Wu L, Maoui M, Wang S, et al. (2002) PTEN associates with the vault particles in HeLa cells. J Biol Chem 277: 40247–40252.
[36]
Ryu SJ, Park SC (2009) Targeting major vault protein in senescence-associated apoptosis resistance. Expert Opin Ther Targets 13: 479–484.
[37]
Mossink MH, de Groot J, van Zon A, Franzel-Luiten E, Schoester M, et al. (2003) Unimpaired dendritic cell functions in MVP/LRP knockout mice. Immunology 110: 58–65.
[38]
Mossink MH, van Zon A, Franzel-Luiten E, Schoester M, Kickhoefer VA, et al. (2002) Disruption of the murine major vault protein (MVP/LRP) gene does not induce hypersensitivity to cytostatics. Cancer Res 62: 7298–7304.
[39]
Glaser P, Frangeul L, Buchrieser C, Rusniok C, Amend A, et al. (2001) Comparative genomics of Listeria species. Science 294: 849–852.
[40]
Bierne H, Sabet C, Personnic N, Cossart P (2007) Internalins: a complex family of leucine-rich repeat-containing proteins in Listeria monocytogenes. Microbes Infect 9: 1156–1166.
[41]
Bierne H, Mazmanian SK, Trost M, Pucciarelli MG, Liu G, et al. (2002) Inactivation of the srtA gene in Listeria monocytogenes inhibits anchoring of surface proteins and affects virulence. Mol Microbiol 43: 869–881.
[42]
Bonazzi M, Cossart P (2006) Bacterial entry into cells: a role for the endocytic machinery. FEBS Lett 580: 2962–2967.
[43]
Camejo A, Buchrieser C, Couve E, Carvalho F, Reis O, et al. (2009) In vivo transcriptional profiling of Listeria monocytogenes and mutagenesis identify new virulence factors involved in infection. PLoS Pathog 5: e1000449.
[44]
Toledo-Arana A, Dussurget O, Nikitas G, Sesto N, Guet-Revillet H, et al. (2009) The Listeria transcriptional landscape from saprophytism to virulence. Nature 459: 950–956.
[45]
Aubry C, Goulard C, Nahori MA, Decalf J, Boneca IG, et al. (2011) OatA, a peptidoglycan O-acetyltransferase involved in Listeria monocytogenes immune escape and critical for virulence. J Infect Dis. Accepted.
[46]
Pucciarelli MG, Calvo E, Sabet C, Bierne H, Cossart P, et al. (2005) Identification of substrates of the Listeria monocytogenes sortases A and B by a non-gel proteomic analysis. Proteomics 5: 4808–4817.
[47]
McGann P, Raengpradub S, Ivanek R, Wiedmann M, Boor KJ (2008) Differential regulation of Listeria monocytogenes internalin and internalin-like genes by sigmaB and PrfA as revealed by subgenomic microarray analyses. Foodborne Pathog Dis 5: 417–435.
[48]
Balestrino D, Hamon MA, Dortet L, Nahori MA, Pizarro-Cerda J, et al. (2010) Single-cell techniques using chromosomally tagged fluorescent bacteria to study Listeria monocytogenes infection processes. Appl Environ Microbiol 76: 3625–3636.
[49]
Sabet C, Toledo-Arana A, Personnic N, Lecuit M, Dubrac S, et al. (2008) The Listeria monocytogenes virulence factor InlJ is specifically expressed in vivo and behaves as an adhesin. Infect Immun 76: 1368–1378.
[50]
Bron PA, Monk IR, Corr SC, Hill C, Gahan CG (2006) Novel luciferase reporter system for in vitro and organ-specific monitoring of differential gene expression in Listeria monocytogenes. Appl Environ Microbiol 72: 2876–2884.
[51]
Kickhoefer VA, Vasu SK, Rome LH (1996) Vaults are the answer, what is the question? Trends Cell Biol 6: 174–178.
[52]
van Zon A, Mossink MH, Scheper RJ, Sonneveld P, Wiemer EA (2003) The vault complex. Cell Mol Life Sci 60: 1828–1837.
[53]
Henry R, Shaughnessy L, Loessner MJ, Alberti-Segui C, Higgins DE, et al. (2006) Cytolysin-dependent delay of vacuole maturation in macrophages infected with Listeria monocytogenes. Cell Microbiol 8: 107–119.
[54]
Mizushima N, Yoshimori T, Levine B (2010) Methods in mammalian autophagy research. Cell 140: 313–326.
[55]
Pankiv S, Clausen TH, Lamark T, Brech A, Bruun JA, et al. (2007) p62/SQSTM1 binds directly to Atg8/LC3 to facilitate degradation of ubiquitinated protein aggregates by autophagy. J Biol Chem 282: 24131–24145.
[56]
Levine B, Mizushima N, Virgin HW (2011) Autophagy in immunity and inflammation. Nature 469: 323–335.
[57]
Izquierdo MA, Scheffer GL, Flens MJ, Giaccone G, Broxterman HJ, et al. (1996) Broad distribution of the multidrug resistance-related vault lung resistance protein in normal human tissues and tumors. Am J Pathol 148: 877–887.
[58]
Sunnaram BL, Gandemer V, Sebillot M, Grandgirard N, Amiot L, et al. (2003) LRP overexpression in monocytic lineage. Leuk Res 27: 755–759.
[59]
Cossart P, Toledo-Arana A (2008) Listeria monocytogenes, a unique model in infection biology: an overview. Microbes Infect 10: 1041–1050.
[60]
Lebreton A, Lakisic G, Job V, Fritsch L, Tham TN, et al. (2011) A Bacterial Protein Targets the BAHD1 Chromatin Complex to Stimulate Type III Interferon Response. Science 331: 1319–1321.
[61]
Kedersha NL, Rome LH (1986) Isolation and characterization of a novel ribonucleoprotein particle: large structures contain a single species of small RNA. J Cell Biol 103: 699–709.
[62]
Persson H, Kvist A, Vallon-Christersson J, Medstrand P, Borg A, et al. (2009) The non-coding RNA of the multidrug resistance-linked vault particle encodes multiple regulatory small RNAs. Nat Cell Biol 11: 1268–1271.
[63]
Suprenant KA (2002) Vault ribonucleoprotein particles: sarcophagi, gondolas, or safety deposit boxes? Biochemistry 41: 14447–14454.
[64]
Nandy C, Mrazek J, Stoiber H, Grasser FA, Huttenhofer A, et al. (2009) Epstein-barr virus-induced expression of a novel human vault RNA. J Mol Biol 388: 776–784.
[65]
Sakaki Y, Terashi K, Yamaguchi A, Kawamata N, Tokito Y, et al. (2002) Human T-cell lymphotropic virus type I Tax activates lung resistance-related protein expression in leukemic clones established from an adult T-cell leukemia patient. Exp Hematol 30: 340–345.
[66]
Ogawa M, Yoshikawa Y, Mimuro H, Hain T, Chakraborty T, et al. (2011) Autophagy targeting of Listeria monocytogenes and the bacterial countermeasure. Autophagy 7: 310–314.
[67]
Arnaud M, Chastanet A, Debarbouille M (2004) New vector for efficient allelic replacement in naturally nontransformable, low-GC-content, gram-positive bacteria. Appl Environ Microbiol 70: 6887–6891.
[68]
Boujemaa-Paterski R, Gouin E, Hansen G, Samarin S, Le Clainche C, et al. (2001) Listeria protein ActA mimics WASp family proteins: it activates filament barbed end branching by Arp2/3 complex. Biochemistry 40: 11390–11404.
[69]
Veiga E, Guttman JA, Bonazzi M, Boucrot E, Toledo-Arana A, et al. (2007) Invasive and adherent bacterial pathogens co-Opt host clathrin for infection. Cell Host Microbe 2: 340–351.
[70]
van Zon A, Mossink MH, Schoester M, Houtsmuller AB, Scheffer GL, et al. (2003) The formation of vault-tubes: a dynamic interaction between vaults and vault PARP. J Cell Sci 116: 4391–4400.
[71]
Mostowy S, Bonazzi M, Hamon MA, Tham TN, Mallet A, et al. (2010) Entrapment of intracytosolic bacteria by septin cage-like structures. Cell Host Microbe 8: 433–444.
[72]
Gouin E, Mengaud J, Cossart P (1994) The virulence gene cluster of Listeria monocytogenes is also present in Listeria ivanovii, an animal pathogen, and Listeria seeligeri, a nonpathogenic species. Infect Immun 62: 3550–3553.
[73]
Levraud JP, Disson O, Kissa K, Bonne I, Cossart P, et al. (2009) Real-time observation of Listeria monocytogenes-phagocyte interactions in living zebrafish larvae. Infect Immun 77: 3651–3660.