Cell-associated hemolysis activity in the clinical strain of Pseudomonas fluorescens MFN1032.
BACKGROUND
MFN1032 is a clinical Pseudomonas fluorescens strain able to grow at 37 degrees C. MFN1032 cells induce necrosis and apoptosis in rat glial cells at this temperature. This strain displays secretion-mediated hemolytic activity involving phospholipase C and cyclolipopeptides. Under laboratory conditions, this activity is not expressed at 37 degrees C. This activity is tightly regulated and is subject to phase variation.


RESULTS
We found that MFN1032 displays a cell-associated hemolytic activity distinct from the secreted hemolytic activity. Cell-associated hemolysis was expressed at 37 degrees C and was only detected in vitro in mid log growth phase in the presence of erythrocytes. We studied the regulation of this activity in the wild-type strain and in a mutant defective in the Gac two-component pathway. GacS/GacA is a negative regulator of this activity. In contrast to the Pseudomonas fluorescens strains PfO-1 and Pf5, whose genomes have been sequenced, the MFN1032 strain has the type III secretion-like genes hrcRST belonging to the hrpU operon. We showed that disruption of this operon abolished cell-associated hemolytic activity. This activity was not detected in P.fluorescens strains carrying similar hrc genes, as for the P. fluorescens psychrotrophic strain MF37.


CONCLUSIONS
To our knowledge this the first demonstration of cell-associated hemolytic activity of a clinical strain of Pseudomonas fluorescens. Moreover, this activity seems to be related to a functional hrpU operon and is independent of biosurfactant production. Precise link between a functional hrpU operon and cell-associated hemolytic activity remains to be elucidated.
Sperandio
Daniel
D
Laboratory of cold microbiology signals and the microenvironment, LMDF-SME, UPRES EA 4312, University of Rouen, 55 rue Saint Germain, 27000 Evreux, France.
Rossignol
Gaelle
G
Guerillon
Josette
J
Connil
Nathalie
N
Orange
Nicole
N
Feuilloley
Marc G J
MG
Merieau
Annabelle
A
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004912:Erythrocytes / Q000382:microbiology; D055786:Gene Knockout Techniques; D006461:Hemolysis; D006801:Humans; D008969:Molecular Sequence Data; D009876:Operon; D011552:Pseudomonas Infections / Q000382:microbiology; D011551:Pseudomonas fluorescens / Q000302:isolation & purification / Q000472:pathogenicity*; D017422:Sequence Analysis, DNA; D012756:Sheep; D013696:Temperature
D001426:Bacterial Proteins; D004269:DNA, Bacterial