Previous Article | Next Article 
Infection and Immunity, October 1998, p. 4602-4610, Vol. 66, No. 10
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Surface-Associated Hsp60 Chaperonin of Legionella
pneumophila Mediates Invasion in a HeLa Cell Model
Rafael A.
Garduño,
Elizabeth
Garduño, and
Paul S.
Hoffman*
Department of Microbiology and Immunology and
Department of Medicine, Division of Infectious Diseases, Dalhousie
University, Halifax, Nova Scotia, Canada B3H 4H7
Received 5 May 1998/Returned for modification 17 June 1998/Accepted 17 July 1998
HeLa cells have been previously used to demonstrate that virulent
strains of Legionella pneumophila (but not salt-tolerant avirulent strains) efficiently invade nonphagocytic cells. Hsp60, a
member of the GroEL family of chaperonins, is displayed on the surface
of virulent L. pneumophila (R. A. Garduño et
al., J. Bacteriol. 180:505-513, 1988). Because Hsp60 is largely
involved in protein-protein interactions, we investigated its role in
adherence-invasion in the HeLa cell model. Hsp60-specific antibodies
inhibited the adherence and invasiveness of two virulent L. pneumophila strains in a dose-dependent manner but had no effect
on the association of their salt-tolerant avirulent derivatives with
HeLa cells. A monospecific anti-OmpS (major outer membrane protein)
serum inhibited the association of both virulent and avirulent strains of L. pneumophila to HeLa cells, suggesting
that while both Hsp60 and OmpS may mediate bacterial association to
HeLa cells, only virulent strains selectively displayed Hsp60 on their
surfaces. Furthermore, the surface-associated Hsp60 of virulent
bacterial cells was susceptible to the action of trypsin, which
rendered the bacteria noninvasive. Additionally, pretreatment of HeLa
cells with purified Hsp60 or precoating of the plastic surface where HeLa cells attached with Hsp60 reduced the adherence and invasiveness of the two virulent strains. Finally, recombinant Hsp60 covalently bound to latex beads promoted the early association of beads with HeLa
cells by a factor of 20 over bovine serum albumin (BSA)-coated beads
and competed with virulent strains for association with HeLa cells.
Hsp60-coated beads were internalized in large numbers by HeLa cells and
remained in tight endosomes that did not fuse with other vesicles,
whereas internalized BSA-coated beads, for which endocytic trafficking
is well established, resided in more loose or elongated endosomes.
Mature intracellular forms of L. pneumophila, which were up
to 100-fold more efficient than agar-grown bacteria at associating with
HeLa cells, were enriched for Hsp60 on the bacterial surface, as
determined by immunolocalization techniques. Collectively, these
results establish a role for surface-exposed Hsp60 in invasion of HeLa
cells by L. pneumophila.
*
Corresponding author. Mailing address: Department of
Microbiology and Immunology, Sir Charles Tupper Medical Building,
Dalhousie University, Halifax, Nova Scotia, Canada B3H 4H7. Phone:
(902) 494-3889. Fax: (902) 494-5125. E-mail:
hoffmanp{at}tupdean1.med.dal.ca.
Infection and Immunity, October 1998, p. 4602-4610, Vol. 66, No. 10
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
Chong, A., Lima, C. A., Allan, D. S., Nasrallah, G. K., Garduno, R. A.
(2009). The Purified and Recombinant Legionella pneumophila Chaperonin Alters Mitochondrial Trafficking and Microfilament Organization. Infect. Immun.
77: 4724-4739
[Abstract]
[Full Text]
-
Morash, M. G., Brassinga, A. K. C., Warthan, M., Gourabathini, P., Garduno, R. A., Goodman, S. D., Hoffman, P. S.
(2009). Reciprocal Expression of Integration Host Factor and HU in the Developmental Cycle and Infectivity of Legionella pneumophila. Appl. Environ. Microbiol.
75: 1826-1837
[Abstract]
[Full Text]
-
Al-Khodor, S., Kalachikov, S., Morozova, I., Price, C. T., Abu Kwaik, Y.
(2009). The PmrA/PmrB Two-Component System of Legionella pneumophila Is a Global Regulator Required for Intracellular Replication within Macrophages and Protozoa. Infect. Immun.
77: 374-386
[Abstract]
[Full Text]
-
Faulkner, G., Berk, S. G., Garduno, E., Ortiz-Jimenez, M. A., Garduno, R. A.
(2008). Passage through Tetrahymena tropicalis Triggers a Rapid Morphological Differentiation in Legionella pneumophila. J. Bacteriol.
190: 7728-7738
[Abstract]
[Full Text]
-
Wuppermann, F. N., Molleken, K., Julien, M., Jantos, C. A., Hegemann, J. H.
(2008). Chlamydia pneumoniae GroEL1 Protein Is Cell Surface Associated and Required for Infection of HEp-2 Cells. J. Bacteriol.
190: 3757-3767
[Abstract]
[Full Text]
-
Galka, F., Wai, S. N., Kusch, H., Engelmann, S., Hecker, M., Schmeck, B., Hippenstiel, S., Uhlin, B. E., Steinert, M.
(2008). Proteomic Characterization of the Whole Secretome of Legionella pneumophila and Functional Analysis of Outer Membrane Vesicles. Infect. Immun.
76: 1825-1836
[Abstract]
[Full Text]
-
Leclerque, A., Kleespies, R. G.
(2008). 16S rRNA-, GroEL- and MucZ-based assessment of the taxonomic position of 'Rickettsiella melolonthae' and its implications for the organization of the genus Rickettsiella. Int. J. Syst. Evol. Microbiol.
58: 749-755
[Abstract]
[Full Text]
-
Berk, S. G., Faulkner, G., Garduno, E., Joy, M. C., Ortiz-Jimenez, M. A., Garduno, R. A.
(2008). Packaging of Live Legionella pneumophila into Pellets Expelled by Tetrahymena spp. Does Not Require Bacterial Replication and Depends on a Dot/Icm-Mediated Survival Mechanism. Appl. Environ. Microbiol.
74: 2187-2199
[Abstract]
[Full Text]
-
Aly, K. A., Krall, L., Lottspeich, F., Baron, C.
(2008). The Type IV Secretion System Component VirB5 Binds to the trans-Zeatin Biosynthetic Enzyme Tzs and Enables Its Translocation to the Cell Surface of Agrobacterium tumefaciens. J. Bacteriol.
190: 1595-1604
[Abstract]
[Full Text]
-
Newton, H. J., Sansom, F. M., Dao, J., McAlister, A. D., Sloan, J., Cianciotto, N. P., Hartland, E. L.
(2007). Sel1 Repeat Protein LpnE Is a Legionella pneumophila Virulence Determinant That Influences Vacuolar Trafficking. Infect. Immun.
75: 5575-5585
[Abstract]
[Full Text]
-
Chatfield, C. H., Cianciotto, N. P.
(2007). The Secreted Pyomelanin Pigment of Legionella pneumophila Confers Ferric Reductase Activity. Infect. Immun.
75: 4062-4070
[Abstract]
[Full Text]
-
Sundaresh, S., Randall, A., Unal, B., Petersen, J. M., Belisle, J. T., Gill Hartley, M., Duffield, M., Titball, R. W., Davies, D. H., Felgner, P. L., Baldi, P.
(2007). From protein microarrays to diagnostic antigen discovery: a study of the pathogen Francisella tularensis. Bioinformatics
23: i508-i518
[Abstract]
[Full Text]
-
Tsugawa, H., Ito, H., Ohshima, M., Okawa, Y.
(2007). Cell adherence-promoted activity of Plesiomonas shigelloides GroEL. J Med Microbiol
56: 23-29
[Abstract]
[Full Text]
-
Huang, B., Yuan, Z., Heron, B. A., Gray, B. R., Eglezos, S., Bates, J. R., Savill, J.
(2006). Distribution of 19 major virulence genes in Legionella pneumophila serogroup 1 isolates from patients and water in Queensland, Australia.. J Med Microbiol
55: 993-997
[Abstract]
[Full Text]
-
Henderson, B., Allan, E., Coates, A. R. M.
(2006). Stress Wars: the Direct Role of Host and Bacterial Molecular Chaperones in Bacterial Infection. Infect. Immun.
74: 3693-3706
[Full Text]
-
Fontes, P., Alvarez-Martinez, M.-T., Gross, A., Carnaud, C., Kohler, S., Liautard, J.-P.
(2005). Absence of Evidence for the Participation of the Macrophage Cellular Prion Protein in Infection with Brucella suis. Infect. Immun.
73: 6229-6236
[Abstract]
[Full Text]
-
Yoshizawa, S., Tateda, K., Matsumoto, T., Gondaira, F., Miyazaki, S., Standiford, T. J., Yamaguchi, K.
(2005). Legionella pneumophila Evades Gamma Interferon-Mediated Growth Suppression through Interleukin-10 Induction in Bone Marrow-Derived Macrophages. Infect. Immun.
73: 2709-2717
[Abstract]
[Full Text]
-
Garmory, H. S., Titball, R. W.
(2004). ATP-Binding Cassette Transporters Are Targets for the Development of Antibacterial Vaccines and Therapies. Infect. Immun.
72: 6757-6763
[Full Text]
-
Grabiec, A., Meng, G., Fichte, S., Bessler, W., Wagner, H., Kirschning, C. J.
(2004). Human but Not Murine Toll-like Receptor 2 Discriminates between Tri-palmitoylated and Tri-lauroylated Peptides. J. Biol. Chem.
279: 48004-48012
[Abstract]
[Full Text]
-
Fischer, S. F., Vier, J., Kirschnek, S., Klos, A., Hess, S., Ying, S., Hacker, G.
(2004). Chlamydia Inhibit Host Cell Apoptosis by Degradation of Proapoptotic BH3-only Proteins. JEM
200: 905-916
[Abstract]
[Full Text]
-
Hartley, M. G., Green, M., Choules, G., Rogers, D., Rees, D. G. C., Newstead, S., Sjostedt, A., Titball, R. W.
(2004). Protection Afforded by Heat Shock Protein 60 from Francisella tularensis Is Due to Copurified Lipopolysaccharide. Infect. Immun.
72: 4109-4113
[Abstract]
[Full Text]
-
Zhang, L., Pelech, S., Uitto, V.-J.
(2004). Long-Term Effect of Heat Shock Protein 60 from Actinobacillus actinomycetemcomitans on Epithelial Cell Viability and Mitogen-Activated Protein Kinases. Infect. Immun.
72: 38-45
[Abstract]
[Full Text]
-
Goulhen, F., Grenier, D., Mayrand, D.
(2003). ORAL MICROBIAL HEAT-SHOCK PROTEINS AND THEIR POTENTIAL CONTRIBUTIONS TO INFECTIONS. CROBM
14: 399-412
[Abstract]
[Full Text]
-
Brassinga, A. K. C., Hiltz, M. F., Sisson, G. R., Morash, M. G., Hill, N., Garduno, E., Edelstein, P. H., Garduno, R. A., Hoffman, P. S.
(2003). A 65-Kilobase Pathogenicity Island Is Unique to Philadelphia-1 Strains of Legionella pneumophila. J. Bacteriol.
185: 4630-4637
[Abstract]
[Full Text]
-
Watarai, M., Kim, S., Erdenebaatar, J., Makino, S.-i., Horiuchi, M., Shirahata, T., Sakaguchi, S., Katamine, S.
(2003). Cellular Prion Protein Promotes Brucella Infection into Macrophages. JEM
198: 5-17
[Abstract]
[Full Text]
-
Belyi, I., Popoff, M. R., Cianciotto, N. P.
(2003). Purification and Characterization of a UDP-Glucosyltransferase Produced by Legionella pneumophila. Infect. Immun.
71: 181-186
[Abstract]
[Full Text]
-
Long, K. H., Gomez, F. J., Morris, R. E., Newman, S. L.
(2003). Identification of Heat Shock Protein 60 as the Ligand on Histoplasma capsulatum That Mediates Binding to CD18 Receptors on Human Macrophages. J. Immunol.
170: 487-494
[Abstract]
[Full Text]
-
Faulkner, G., Garduno, R. A.
(2002). Ultrastructural Analysis of Differentiation in Legionella pneumophila. J. Bacteriol.
184: 7025-7041
[Abstract]
[Full Text]
-
Garduno, R. A., Garduno, E., Hiltz, M., Hoffman, P. S.
(2002). Intracellular Growth of Legionella pneumophila Gives Rise to a Differentiated Form Dissimilar to Stationary-Phase Forms. Infect. Immun.
70: 6273-6283
[Abstract]
[Full Text]
-
Dietrich, C., Heuner, K., Brand, B. C., Hacker, J., Steinert, M.
(2001). Flagellum of Legionella pneumophila Positively Affects the Early Phase of Infection of Eukaryotic Host Cells. Infect. Immun.
69: 2116-2122
[Abstract]
[Full Text]
-
Hennequin, C., Porcheray, F., Waligora-Dupriet, A.-J., Collignon, A., Barc, M.-C., Bourlioux, P., Karjalainen, T.
(2001). GroEL (Hsp60) of Clostridium difficile is involved in cell adherence. Microbiology
147: 87-96
[Abstract]
[Full Text]
-
Resto-Ruiz, S. I., Sweger, D., Widen, R. H., Valkov, N., Anderson, B. E.
(2000). Transcriptional Activation of the htrA (High-Temperature Requirement A) Gene from Bartonella henselae. Infect. Immun.
68: 5970-5978
[Abstract]
[Full Text]
-
Zaborina, O., Misra, N., Kostal, J., Kamath, S., Kapatral, V., El-Idrissi, M. E.-A., Prabhakar, B. S., Chakrabarty, A. M.
(1999). P2Z-Independent and P2Z Receptor-Mediated Macrophage Killing by Pseudomonas aeruginosa Isolated from Cystic Fibrosis Patients. Infect. Immun.
67: 5231-5242
[Abstract]
[Full Text]
-
Gao, L.-Y., Abu Kwaik, Y.
(1999). Activation of Caspase 3 during Legionella pneumophila-Induced Apoptosis. Infect. Immun.
67: 4886-4894
[Abstract]
[Full Text]