The ability of Legionella pneumophila (Lpn) to cause pneumonia is dependent on intracellular replication within pulmonary phagocytic and epithelial cells. In the environment, the bacteria are ubiquitous where they multiply intracellularly with amoebae. Invasion and intracellular replication within protozoa play major factors in the amplification and dissemination of Lpn in the environment and in transmission and infectivity to humans. We have recently shown that uptake of Lpn by the protozoan Hartmannella vermiformis is mediated by bacterial attachment to a b2 integrin-like Galactose/N-acetyl-galactosamine lectin on the protozoan surface. Bacterial attachment to the lectin is associated with induction of protozoan gene expression and with tyrosine dephosphorylation of the lectin and several cytoskeletal proteins including actin, focal adhesion kinase, paxillin, and vinculin. Our data suggested a cytoskeletal disruption in the protozoan host upon bacterial attachment. Transmission electron microscopy showed that these bacterial-induced manipulations of cell processes in the protozoan host are associated with entry of the majority of the bacteria through a cup shape-like invagination that resemble receptor-mediated endocytosis, but some bacteria are internalized by coiling phagocytosis. Our preliminary data suggest that the mechanism of entry of Lpn is novel, which may contribute to its subsequent intracellular fate. Our hypothesis is that the lectin is a protozoan receptor involved in uptake of Lpn, and is dissociated from the cytoskeleton upon bacterial attachment and invasion.
Our specific aims are, 1) to clone the lectin encoding gene and examine its regulation of expression; 2) to clone and characterize the bacterial ligand that binds the lectin receptor and the mode of ligand-receptor interaction; 3) to determine cellular distribution of the lectin receptor and its subsequent fate after internalization; and 4) to evaluate the interaction of the receptor with the cytoskeleton. The results derived for the proposed studies will uncover new paradigms of uptake of intracellular pathogens and will contribute to our understanding of targeting of molecules into a """"""""protected vacuole"""""""" inside eukaryotic cells. Our proposed studies may also facilitate the design of future preventive strategies to control the amplification and spread of Lpn in the aquatic environment, which is the only source of bacterial transmission to humans. Our studies may uncover potential pathogenic evolution of Lpn to invade the more evolved mammalian cells, and may contribute to the understanding of invasion of protozoa by Mycobacterium and Chlamydia.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI043965-03
Application #
6511087
Study Section
Bacteriology and Mycology Subcommittee 2 (BM)
Program Officer
Korpela, Jukka K
Project Start
2000-07-15
Project End
2005-05-31
Budget Start
2002-07-01
Budget End
2003-05-31
Support Year
3
Fiscal Year
2002
Total Cost
$240,345
Indirect Cost
Name
University of Kentucky
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
832127323
City
Lexington
State
KY
Country
United States
Zip Code
40506
Price, Christopher T D; Richards, Ashley M; Von Dwingelo, Juanita E et al. (2014) Amoeba host-Legionella synchronization of amino acid auxotrophy and its role in bacterial adaptation and pathogenic evolution. Environ Microbiol 16:350-8
Abu Kwaik, Yousef; Bumann, Dirk (2013) Microbial quest for food in vivo: 'nutritional virulence' as an emerging paradigm. Cell Microbiol 15:882-90
Al-Quadan, Tasneem; Price, Christopher T; Abu Kwaik, Yousef (2012) Exploitation of evolutionarily conserved amoeba and mammalian processes by Legionella. Trends Microbiol 20:299-306
Akimana, Christine; Kwaik, Yousef Abu (2011) Francisella-arthropod vector interaction and its role in patho-adaptation to infect mammals. Front Microbiol 2:34
Al-Quadan, Tasneem; Price, Christopher T; London, Nir et al. (2011) Anchoring of bacterial effectors to host membranes through host-mediated lipidation by prenylation: a common paradigm. Trends Microbiol 19:573-9
Price, Christopher T D; Al-Quadan, Tasneem; Santic, Marina et al. (2011) Host proteasomal degradation generates amino acids essential for intracellular bacterial growth. Science 334:1553-7
Asare, Rexford; Akimana, Christine; Jones, Snake et al. (2010) Molecular bases of proliferation of Francisella tularensis in arthropod vectors. Environ Microbiol 12:2587-612
Asare, Rexford; Abu Kwaik, Yousef (2010) Molecular complexity orchestrates modulation of phagosome biogenesis and escape to the cytosol of macrophages by Francisella tularensis. Environ Microbiol 12:2559-86
Santic, Marina; Pavokovic, Gordana; Jones, Snake et al. (2010) Regulation of apoptosis and anti-apoptosis signalling by Francisella tularensis. Microbes Infect 12:126-34
Santic, Marina; Al-Khodor, Souhaila; Abu Kwaik, Yousef (2010) Cell biology and molecular ecology of Francisella tularensis. Cell Microbiol 12:129-39

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