Abstract
Francisella tularensis harbors genes with similarity to genes encoding components of a type VI secretion system (T6SS). These include iglA and iglB, the homologues of which are conserved in T6SSs. They are part of the igl operon, also encompassing the iglC and iglD genes. We have used a yeast two-hybrid system to study the interaction of the Igl proteins of F. tularensis LVS. Previously, we identified a region of IglA necessary for efficient binding to IglB as well as for IglAB protein stability and intra-macrophage growth with an essential role for a conserved α-helical region. Thus, IglA-IglB complex formation is clearly crucial for Francisella pathogenicity and the same interaction is conserved in other human pathogens. Herein, the interaction of IglC with other members of the operon was investigated. It showed no binding to the other members in the yeast two-hybrid assay and we found also that two cysteine residues, C191 and C192, predicted to be putative prenylation sites, played no role for the important contribution of IglC to the intracellular replication of F. tularensis although C191 was important for the stability of the protein.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Barker, J. R. & Klose, K. E. (2009) Characterization of the secreted effector protein IglC in Francisella tularensis virulence. The 6th International conference on tularemia, Berlin, Robert Koch Institut.
Bingle, L. E., Bailey, C. M. et al. (2008) Type VI secretion: a beginner’s guide. Curr Opin Microbiol, 11(1), 3–8.
Bröms, J. E., Lavander, M. et al. (2009) A conserved alpha-helix essential for a type VI secretion-like system of Francisella tularensis. J Bacteriol, 191(8), 2431–46.
Bönquist, L., Lindgren, H. et al. (2008) MglA and Igl proteins contribute to the modulation of Francisella tularensis live vaccine strain-containing phagosomes in murine macrophages. Infect Immun, 76(8), 3502–10.
Chamberlain, R. E. (1965) Evaluation of live tularemia vaccine prepared in a chemically defined medium. Appl Microbiol, 13, 232–5.
Das, S. & Chaudhuri, K. (2003) Identification of a unique IAHP (IcmF associated homologous proteins) cluster in Vibrio cholerae and other proteobacteria through in silico analysis. In Silico Biol, 3(3), 287–300.
De Bruin, O. M., Ludu, J. S. et al. (2007) The Francisella pathogenicity island protein IglA localizes to the bacterial cytoplasm and is needed for intracellular growth. BMC Microbiol, 7(1), 1.
Dudley, E. G., Thomson, N. R. et al. (2006) Proteomic and microarray characterization of the AggR regulon identifies a pheU pathogenicity island in enteroaggregative Escherichia coli. Mol Microbiol, 61(5), 1267–82.
Farnsworth, C., Seabra, M. et al. (1994) Rab geranylgeranyl transferase catalyzes the geranylgeranylation of adjacent cysteines in the small GTPases Rab1A, Rab3A, and Rab5A. Proc Natl Acad Sci USA, 91, 11963–7.
Feldman, M. F., Muller, S. et al. (2002) SycE allows secretion of YopE-DHFR hybrids by the Yersinia enterocolitica type III Ysc system. Mol Microbiol, 46(4), 1183–97.
Golovliov, I., Sjöstedt, A. et al. (2003) A method for allelic replacement in Francisella tularensis. FEMS Microbiol Lett, 222(2), 273–80.
James, P., Halladay, J. et al. (1996) Genomic libraries and a host strain designed for highly efficient two-hybrid selection in yeast. Genetics, 144(4), 1425–36.
Kadzhaev, K., Zingmark, C. et al. (2009) Identification of genes contributing to the virulence of Francisella tularensis SCHU S4 in a mouse intradermal infection model. PLoS One, 4(5), e5463.
Lenco, J., Pavkova, I. et al. (2005) Insights into the oxidative stress response in Francisella tularensis LVS and its mutant DeltaiglC1+2 by proteomics analysis. FEMS Microbiol Lett, 246(1), 47–54.
Lindgren, H., Golovliov, I. et al. (2004) Factors affecting the escape of Francisella tularensis from the phagolysosome. J Med Microbiol, 53(Pt 10), 953–8.
Ludu, J. S., De Bruin, O. M. et al. (2008) The Francisella pathogenicity island protein PdpD is required for full virulence and associates with homologues of the type VI secretion system. J Bacteriol, 190(13), 4584–95.
Mougous, J. D., Cuff, M. E. et al. (2006) A virulence locus of Pseudomonas aeruginosa encodes a protein secretion apparatus. Science, 312(5779), 1526–30.
Nano, F. E. & Schmerk, C. (2007) The Francisella pathogenicity island. Ann NY Acad Sci, 1105, 122–37.
Nano, F. E., Zhang, N. et al. (2004) A Francisella tularensis Pathogenicity Island Required for Intramacrophage Growth. J Bacteriol, 186(19), 6430–6.
Pereira-Leal, J. & Seabra, M. C. (2001) Evolution of the Rab family of small GTP-binding proteins. J Mol Biol, 313, 889–901.
Pukatzki, S., Ma, A. T. et al. (2006) Identification of a conserved bacterial protein secretion system in Vibrio cholerae using the Dictyostelium host model system. Proc Natl Acad Sci USA, 103(5), 1528–33.
Santic, M., Molmeret, M. et al. (2007) A Francisella tularensis pathogenicity island protein essential for bacterial proliferation within the host cell cytosol. Cell Microbiol, 9(10), 2391–403.
Schell, M. A., Ulrich, R. L. et al. (2007) Type VI secretion is a major virulence determinant in Burkholderia mallei. Mol Microbiol, 64(6), 1466–85.
Suarez, G., Sierra, J. C. et al. (2008) Molecular characterization of a functional type VI secretion system from a clinical isolate of Aeromonas hydrophila. Microb Pathog, 44(4), 344–61.
Twine, S., Bystrom, M. et al. (2005) A mutant of Francisella tularensis strain SCHU S4 lacking the ability to express a 58-kilodalton protein is attenuated for virulence and is an effective live vaccine. Infect Immun, 73(12), 8345–52.
Zheng, J. & Leung, K. Y. (2007) Dissection of a type VI secretion system in Edwardsiella tarda. Mol Microbiol, 66(5), 1192–206.
Acknowledgements
This work was supported by grants 2006–3426 (JB) and 2006–2877 (AS) from the Swedish Research Council and a grant from the Medical Faculty, Umeå University, and the Umeå Biotech Incubator, Umeå, Sweden. The work was performed in part at the Umeå Centre for Microbial Research (UCMR).
Index words: Francisella, type VI secretion system, Igl proteins, yeast two-hybrid system, IglC, cysteine residues.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Springer Science+Business Media B.V.
About this paper
Cite this paper
Bröms, J.E., Lavander, M., Sjöstedt, A. (2010). Dissection of the Functions of the IglC Protein of Francisella tularensis . In: Shafferman, A., Ordentlich, A., Velan, B. (eds) The Challenge of Highly Pathogenic Microorganisms. Springer, Dordrecht. https://doi.org/10.1007/978-90-481-9054-6_7
Download citation
DOI: https://doi.org/10.1007/978-90-481-9054-6_7
Published:
Publisher Name: Springer, Dordrecht
Print ISBN: 978-90-481-9053-9
Online ISBN: 978-90-481-9054-6
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)