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PMID: 9009317 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Simultaneous prevention of glutamine synthesis and high-affinity transport attenuates Salmonella typhimurium virulence.

Infection and immunity ·Vol. 65 ·No. 2 ·1997-02-00 ·Pages 587-96

Klose KE, Mekalanos JJ

Abstract

In Salmonella typhimurium, transcription of the glnA gene (encoding glutamine synthetase) is under the control of the nitrogen-regulatory (ntr) system comprising the alternate sigma factor sigma54 (NtrA) and the two-component sensor-transcriptional activator pair NtrB and NtrC. The glnA, ntrB, and ntrC genes form an operon. We measured the virulence of S. typhimurium strains with nitrogen-regulatory mutations after intraperitoneal (i.p.) or oral inoculations of BALB/c mice. Strains with single mutations in glnA, ntrA, ntrB, or ntrC had i.p. 50% lethal doses (LD50s) of <10 bacteria, similar to the wild-type strain. However, a strain with a delta(glnA-ntrC) operon deletion had an i.p. LD50 of >10(5) bacteria, as did delta glnA ntrA and delta glnA ntrC strains, suggesting that glnA strains require an ntr-transcribed gene for full virulence. High-level transcription of the glutamine transport operon (glnHPQ) is dependent upon both ntrA and ntrC, as determined by glnHp-lacZ fusion measurements. Moreover, delta glnA glnH and delta glnA glnQ strains are attenuated, similar to delta glnA ntrA and delta glnA ntrC strains. These results reveal that access of S. typhimurium to host glutamine depends on the ntr system, which apparently is required for the transcription of the glutamine transport genes. The delta(glnA-ntrC) strain exhibited a reduced ability to survive within the macrophage cell line J774, identifying a potential host environment with low levels of glutamine. Finally, the delta(glnA-ntrC) strain, when inoculated at doses as low as 10 organisms, provided mice with protective immunity against challenge by the wild-type strain, demonstrating its potential use as a live vaccine.

MeSH Terms
Animals Base Sequence Biological Transport/immunology Female Gene Expression Regulation, Bacterial/immunology Genes, Bacterial Glutamine/antagonists & inhibitors,deficiency,metabolism Macrophages/microbiology Mice Mice, Inbred BALB C Molecular Sequence Data Mutagenesis, Insertional Mutation Nitrogen/physiology Operon Salmonella typhimurium/genetics,growth & development,pathogenicity Transcription, Genetic Virulence
Chemicals
Glutamine Nitrogen
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Klose K E
Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA.
Mekalanos J J
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Article Info
Journal
Infection and immunity
Abbr.
Infect Immun
ISSN
0019-9567
DOI
10.1128/iai.65.2.587-596.1997
Published
1997-02-00
Pages
587-96
Language
English
Region
United States
NLM ID
0246127
PMCID
PMC176100
Subset
IM
Grants
NIAID NIH HHS · AI09118-03 · United States
NIAID NIH HHS · AI26289 · United States
Databases
GENBANK
U73111
External Links
PubMed source DOI: 10.1128/iai.65.2.587-596.1997 PMC Full Text View journal details
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