Mutation in tcpR gene (Vc0832) of Vibrio cholerae O1 causes loss of tolerance to high osmolarity and affects colonization and virulence in infant mice.
Vibrio cholerae, the agent of cholera, multiplies and colonizes human intestinal tract where it survives high osmolarity due to bile and other sodium salts. In this work, by TnphoA mutagenesis, a mutant of V. cholerae O1 which could not grow and form colonies on LB agar containing 400 mM NaCl has been characterized. The mutant, designated CD83, adhered normally to freshly isolated rabbit intestinal discs, colonized poorly the gut of infant mice and was avirulent in the same model, whereas the parent strain CD81 would colonize the gut and cause death of mice. Attenuation of virulence of CD83 was not attributable to its inability to produce cholera toxin, as no difference was found in the cholera toxin produced in vitro by the mutant and parent strains. Molecular cloning and sequencing of the mutated gene revealed that insertion of transposon occurred in tcpR gene (Vc0832) of V. cholerae. Complementation of the mutant with wild-type tcpR gene resulted in restoration of the ability to survive at high salt concentration (400 mM NaCl), and to colonize the gut and restore virulence. The results suggest that tcpR plays a role in survival of V. cholerae in the small intestine of host as the osmolarity in the intestinal lumen is thought to be equivalent to 300 mM NaCl or higher.
Mishra
Arunima
A
Microbiology Division, Central Drug Research Institute, Lucknow 226001, India 2Institute of Microbiology, University of Ancona, Ancona, Italy.
Srivastava
Ranjana
R
Microbiology Division, Central Drug Research Institute, Lucknow 226001, India 2Institute of Microbiology, University of Ancona, Ancona, Italy.
Pruzzo
Carla
C
Microbiology Division, Central Drug Research Institute, Lucknow 226001, India 2Institute of Microbiology, University of Ancona, Ancona, Italy.
Srivastava
Brahm S
BS
Microbiology Division, Central Drug Research Institute, Lucknow 226001, India 2Institute of Microbiology, University of Ancona, Ancona, Italy.
D000818:Animals; D003001:Cloning, Molecular; D010861:Fimbriae, Bacterial / Q000235:genetics*; D051379:Mice; D009154:Mutation; D009876:Operon; D009994:Osmolar Concentration; D016133:Polymerase Chain Reaction; D012965:Sodium Chloride / Q000494:pharmacology; D014734:Vibrio cholerae / Q000235:genetics / Q000472:pathogenicity*; D014774:Virulence; D037521:Virulence Factors / Q000235:genetics*
D037521:Virulence Factors; D012965:Sodium Chloride