Multiple deletions of the osmolyte transporters BetL, Gbu, and OpuC of Listeria monocytogenes affect virulence and growth at high osmolarity

Typeset version

 

TY  - JOUR
  - Wemekamp-Kamphuis, HH,Wouters, JA,Sleator, RD,Gahan, CGM,Hill, C,Abee, T
  - 2002
  - February
  - Applied and Environmental Microbiology
  - Multiple deletions of the osmolyte transporters BetL, Gbu, and OpuC of Listeria monocytogenes affect virulence and growth at high osmolarity
  - Validated
  - ()
  - OSMOPROTECTANT GLYCINE BETAINE BACILLUS-SUBTILIS COMPATIBLE SOLUTES SALT TOLERANCE ABC TRANSPORTERS STRESS RESPONSES ESCHERICHIA-COLI MINIMAL MEDIUM UPTAKE SYSTEM IDENTIFICATION
  - 68
  - 4710
  - 4716
  - The success of Listeria monocytogenes as a food-borne pathogen owes much to its ability to survive a variety of stresses, both in the food environment and, after ingestion, within the animal host. Growth at high salt concentrations is attributed mainly to the accumulation of organic solutes such as glycine betaine and carnitine. We characterized L. monocytogenes LO28 strains with single, double, and triple deletions in the osmolyte transport systems BetL, Gbu, and OpuC. When single deletion mutants were tested, Gbu was found to have the most drastic effect on the rate of growth in brain heart infusion (13111) broth with 6% added NaCl. The highest reduction in growth rate was found for the triple mutant LO28BCG (DeltabetL DeltaopuC Deltagbu), although the mutant was still capable of growth under these adverse conditions. In addition, we analyzed the growth and survival of this triple mutant in an animal (murine) model. LO28BCG showed a significant reduction in its ability to cause systemic infection following peroral coinoculation with the wild-type parent. Altering OpuC alone resulted in similar effects (R. D. Sleator, J. Wouters, C. G. M. Gahan, T. Abee, and C. Hill, Appl. Environ. Microbiol. 67:2692-2698, 2001), leading to the assumption that OpuC may play an important role in listerial pathogenesis. Analysis of the accumulation of osmolytes revealed that betaine is accumulated up to 300 mumol/g (dry weight) when grown in 13111 broth plus 6% NaCl whereas no carnitine accumulation could be detected. Radiolabeled-betaine uptake studies revealed an inability of BGSOE (DeltabetL Deltagbu) and LO28BCG to transport betaine. Indeed, for LO28BCG, no accumulated betaine was found, but carnitine was accumulated in this strain up to 600 mumol/g (dry weight) of cells, indicating the presence of a possible fourth osmolyte transporter.
  - DOI 10.1128/AEM.68.10.4710-4716.2002
DA  - 2002/02
ER  - 
@article{V43338072,
   = {Wemekamp-Kamphuis,  HH and Wouters,  JA and Sleator,  RD and Gahan,  CGM and Hill,  C and Abee,  T },
   = {2002},
   = {February},
   = {Applied and Environmental Microbiology},
   = {Multiple deletions of the osmolyte transporters BetL, Gbu, and OpuC of Listeria monocytogenes affect virulence and growth at high osmolarity},
   = {Validated},
   = {()},
   = {OSMOPROTECTANT GLYCINE BETAINE BACILLUS-SUBTILIS COMPATIBLE SOLUTES SALT TOLERANCE ABC TRANSPORTERS STRESS RESPONSES ESCHERICHIA-COLI MINIMAL MEDIUM UPTAKE SYSTEM IDENTIFICATION},
   = {68},
  pages = {4710--4716},
   = {{The success of Listeria monocytogenes as a food-borne pathogen owes much to its ability to survive a variety of stresses, both in the food environment and, after ingestion, within the animal host. Growth at high salt concentrations is attributed mainly to the accumulation of organic solutes such as glycine betaine and carnitine. We characterized L. monocytogenes LO28 strains with single, double, and triple deletions in the osmolyte transport systems BetL, Gbu, and OpuC. When single deletion mutants were tested, Gbu was found to have the most drastic effect on the rate of growth in brain heart infusion (13111) broth with 6% added NaCl. The highest reduction in growth rate was found for the triple mutant LO28BCG (DeltabetL DeltaopuC Deltagbu), although the mutant was still capable of growth under these adverse conditions. In addition, we analyzed the growth and survival of this triple mutant in an animal (murine) model. LO28BCG showed a significant reduction in its ability to cause systemic infection following peroral coinoculation with the wild-type parent. Altering OpuC alone resulted in similar effects (R. D. Sleator, J. Wouters, C. G. M. Gahan, T. Abee, and C. Hill, Appl. Environ. Microbiol. 67:2692-2698, 2001), leading to the assumption that OpuC may play an important role in listerial pathogenesis. Analysis of the accumulation of osmolytes revealed that betaine is accumulated up to 300 mumol/g (dry weight) when grown in 13111 broth plus 6% NaCl whereas no carnitine accumulation could be detected. Radiolabeled-betaine uptake studies revealed an inability of BGSOE (DeltabetL Deltagbu) and LO28BCG to transport betaine. Indeed, for LO28BCG, no accumulated betaine was found, but carnitine was accumulated in this strain up to 600 mumol/g (dry weight) of cells, indicating the presence of a possible fourth osmolyte transporter.}},
   = {DOI 10.1128/AEM.68.10.4710-4716.2002},
  source = {IRIS}
}
AUTHORSWemekamp-Kamphuis, HH,Wouters, JA,Sleator, RD,Gahan, CGM,Hill, C,Abee, T
YEAR2002
MONTHFebruary
JOURNAL_CODEApplied and Environmental Microbiology
TITLEMultiple deletions of the osmolyte transporters BetL, Gbu, and OpuC of Listeria monocytogenes affect virulence and growth at high osmolarity
STATUSValidated
TIMES_CITED()
SEARCH_KEYWORDOSMOPROTECTANT GLYCINE BETAINE BACILLUS-SUBTILIS COMPATIBLE SOLUTES SALT TOLERANCE ABC TRANSPORTERS STRESS RESPONSES ESCHERICHIA-COLI MINIMAL MEDIUM UPTAKE SYSTEM IDENTIFICATION
VOLUME68
ISSUE
START_PAGE4710
END_PAGE4716
ABSTRACTThe success of Listeria monocytogenes as a food-borne pathogen owes much to its ability to survive a variety of stresses, both in the food environment and, after ingestion, within the animal host. Growth at high salt concentrations is attributed mainly to the accumulation of organic solutes such as glycine betaine and carnitine. We characterized L. monocytogenes LO28 strains with single, double, and triple deletions in the osmolyte transport systems BetL, Gbu, and OpuC. When single deletion mutants were tested, Gbu was found to have the most drastic effect on the rate of growth in brain heart infusion (13111) broth with 6% added NaCl. The highest reduction in growth rate was found for the triple mutant LO28BCG (DeltabetL DeltaopuC Deltagbu), although the mutant was still capable of growth under these adverse conditions. In addition, we analyzed the growth and survival of this triple mutant in an animal (murine) model. LO28BCG showed a significant reduction in its ability to cause systemic infection following peroral coinoculation with the wild-type parent. Altering OpuC alone resulted in similar effects (R. D. Sleator, J. Wouters, C. G. M. Gahan, T. Abee, and C. Hill, Appl. Environ. Microbiol. 67:2692-2698, 2001), leading to the assumption that OpuC may play an important role in listerial pathogenesis. Analysis of the accumulation of osmolytes revealed that betaine is accumulated up to 300 mumol/g (dry weight) when grown in 13111 broth plus 6% NaCl whereas no carnitine accumulation could be detected. Radiolabeled-betaine uptake studies revealed an inability of BGSOE (DeltabetL Deltagbu) and LO28BCG to transport betaine. Indeed, for LO28BCG, no accumulated betaine was found, but carnitine was accumulated in this strain up to 600 mumol/g (dry weight) of cells, indicating the presence of a possible fourth osmolyte transporter.
PUBLISHER_LOCATION
ISBN_ISSN
EDITION
URL
DOI_LINKDOI 10.1128/AEM.68.10.4710-4716.2002
FUNDING_BODY
GRANT_DETAILS