The Francisella pathogenicity island protein IglA localizes to the bacterial cytoplasm and is needed for intracellular growth.
BACKGROUND
Francisella tularensis is a gram negative, facultative intracellular bacterium that is the etiological agent of tularemia. F. novicida is closely related to F. tularensis but has low virulence for humans while being highly virulent in mice. IglA is a 21 kDa protein encoded by a gene that is part of an iglABCD operon located on the Francisella pathogenicity island (FPI).


RESULTS
Bioinformatics analysis of the FPI suggests that IglA and IglB are components of a newly described type VI secretion system. In this study, we showed that IglA regulation is controlled by the global regulators MglA and MglB. During intracellular growth IglA production reaches a maximum at about 10 hours post infection. Biochemical fractionation showed that IglA is a soluble cytoplasmic protein and immunoprecipitation experiments demonstrate that it interacts with the downstream-encoded IglB. When the iglB gene was disrupted IglA could not be detected in cell extracts of F. novicida, although IglC could be detected. We further demonstrated that IglA is needed for intracellular growth of F. novicida. A non-polar iglA deletion mutant was defective for growth in mouse macrophage-like cells, and in cis complementation largely restored the wild type macrophage growth phenotype.


CONCLUSIONS
The results of this study demonstrate that IglA and IglB are interacting cytoplasmic proteins that are required for intramacrophage growth. The significance of the interaction may be to secrete effector molecules that affect host cell processes.
de Bruin
Olle M
OM
The Department of Biochemistry and Microbiology, University of Victoria, Victoria, BC, Canada. debruin@uvic.ca <debruin@uvic.ca>
Ludu
Jagjit S
JS
Nano
Francis E
FE
D000818:Animals; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology*; D002458:Cell Fractionation; D019295:Computational Biology; D003593:Cytoplasm / Q000737:chemistry*; D005604:Francisella tularensis / Q000737:chemistry / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity*; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D005816:Genetic Complementation Test; D044404:Genomic Islands / Q000235:genetics; D047468:Immunoprecipitation; D008264:Macrophages / Q000382:microbiology; D051379:Mice; D016254:Mutagenesis, Insertional; D011485:Protein Binding; D021381:Protein Transport / Q000235:genetics
D001426:Bacterial Proteins