Differential expression of the enolase gene under in vivo versus in vitro growth conditions of Aeromonas hydrophila.
Aeromonas hydrophila is an emerging human pathogen that leads to gastroenteritis and other invasive diseases. By using a murine peritoneal culture (MPC) model, we identified via restriction fragment differential display PCR (RFDDPCR) five genes of A. hydrophila that were differentially expressed under in vivo versus in vitro growth conditions. The gene encoding enolase was among those five genes that were differentially up regulated. Enolase is a glycolytic enzyme and its surface expression was recently shown to be important in the pathogenesis of a gram-positive bacterium Streptococcus pyogenes. By Western blot analysis and Immunogold staining, we demonstrated secretion and surface expression of enolase in A. hydrophila. We also showed that the whole cells of A. hydrophila had strong enolase activity. Using an enzyme-linked immunosorbant assay and sandwich Western blot analysis, we demonstrated binding of enolase to human plasminogen, which is involved in the fibrinolytic system of the host. We cloned the A. hydrophila enolase gene, which exhibited 62% homology at the DNA level and 57% homology at the amino acid level when compared to S. pyogenes enolase. This is a first report describing the increased expression of enolase gene in vivo that could potentially contribute to the pathogenesis of A. hydrophila infections.
Sha
Jian
J
Department of Microbiology and Immunology, 301 University Blvd, Medical Research Building, The University of Texas Medical Branch, Galveston, TX 77555-1070, USA.
Galindo
C L
CL
Pancholi
V
V
Popov
V L
VL
Zhao
Y
Y
Houston
C W
CW
Chopra
A K
AK
D016980:Aeromonas hydrophila / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development / Q000472:pathogenicity; D000818:Animals; D004269:DNA, Bacterial / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial; D016905:Gram-Negative Bacterial Infections / Q000209:etiology; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D016253:Microscopy, Immunoelectron; D010751:Phosphopyruvate Hydratase / Q000235:genetics* / Q000378:metabolism; D014774:Virulence / Q000235:genetics / Q000502:physiology
D004269:DNA, Bacterial; D010751:Phosphopyruvate Hydratase