Identification of genes contributing to the virulence of Francisella tularensis SCHU S4 in a mouse intradermal infection model.
BACKGROUND
Francisella tularensis is a highly virulent human pathogen. The most virulent strains belong to subspecies tularensis and these strains cause a sometimes fatal disease. Despite an intense recent research effort, there is very limited information available that explains the unique features of subspecies tularensis strains that distinguish them from other F. tularensis strains and that explain their high virulence. Here we report the use of targeted mutagenesis to investigate the roles of various genes or pathways for the virulence of strain SCHU S4, the type strain of subspecies tularensis.


RESULTS
The virulence of SCHU S4 mutants was assessed by following the outcome of infection after intradermal administration of graded doses of bacteria. By this route, the LD(50) of the SCHU S4 strain is one CFU. The virulence of 20 in-frame deletion mutants and 37 transposon mutants was assessed. A majority of the mutants did not show increased prolonged time to death, among them notably Delta pyrB and Delta recA. Of the remaining, mutations in six unique targets, tolC, rep, FTT0609, FTT1149c, ahpC, and hfq resulted in significantly prolonged time to death and mutations in nine targets, rplA, wbtI, iglB, iglD, purL, purF, ggt, kdtA, and glpX, led to marked attenuation with an LD(50) of > 10(3) CFU. In fact, the latter seven mutants showed very marked attenuation with an LD(50) of > or = 10(7) CFU.


CONCLUSIONS
The results demonstrate that the characterization of targeted mutants yielded important information about essential virulence determinants that will help to identify the so far little understood extreme virulence of F. tularensis subspecies tularensis.
Department of Clinical Microbiology, Clinical Bacteriology, and Laboratory for Molecular Infection Medicine Sweden, Umeå University, Umeå, Sweden.
Konstantin
K
Kadzhaev
Carl
C
Zingmark
Igor
I
Golovliov
Mark
M
Bolanowski
Hua
H
Shen
Wayne
W
Conlan
Anders
A
Sjöstedt
D000818:Animals; D004251:DNA Transposable Elements / Q000235:genetics; D004195:Disease Models, Animal; D005604:Francisella tularensis / Q000235:genetics* / Q000472:pathogenicity*; D005798:Genes, Bacterial; D051379:Mice; D050296:Microbial Viability; D009154:Mutation / Q000235:genetics; D012871:Skin Diseases / Q000150:complications / Q000235:genetics* / Q000382:microbiology*; D014406:Tularemia / Q000150:complications / Q000235:genetics* / Q000382:microbiology*; D014774:Virulence / Q000235:genetics
D004251:DNA Transposable Elements