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Identification and disruption of BetL, a secondary glycine betaine transport system linked to the salt tolerance of Listeria monocytogenes LO28.

ABSTRACT: The trimethylammonium compound glycine betaine (N,N, N-trimethylglycine) can be accumulated to high intracellular concentrations, conferring enhanced osmo- and cryotolerance upon Listeria monocytogenes. We report the identification of betL, a gene encoding a glycine betaine uptake system in L. monocytogenes, isolated by functional complementation of the betaine uptake mutant Escherichia coli MKH13. The betL gene is preceded by a consensus sigmaB-dependent promoter and is predicted to encode a 55-kDa protein (507 amino acid residues) with 12 transmembrane regions. BetL exhibits significant sequence homologies to other glycine betaine transporters, including OpuD from Bacillus subtilis (57% identity) and BetP from Corynebacterium glutamicum (41% identity). These high-affinity secondary transporters form a subset of the trimethylammonium transporter family specific for glycine betaine, whose substrates possess a fully methylated quaternary ammonium group. The observed Km value of 7.9 microM for glycine betaine uptake after heterologous expression of betL in E. coli MKH13 is consistent with values obtained for L. monocytogenes in other studies. In addition, a betL knockout mutant which is significantly affected in its ability to accumulate glycine betaine in the presence or absence of NaCl has been constructed in L. monocytogenes. This mutant is also unable to withstand concentrations of salt as high as can the BetL+ parent, signifying the role of the transporter in Listeria osmotolerance.

SUBMITTER: Sleator RD

PROVIDER: S-EPMC91301 | biostudies-literature | 1999 May

REPOSITORIES: biostudies-literature

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Identification and disruption of BetL, a secondary glycine betaine transport system linked to the salt tolerance of Listeria monocytogenes LO28.

Sleator R D RD Gahan C G CG Abee T T Hill C C

Applied and environmental microbiology 19990501 5

The trimethylammonium compound glycine betaine (N,N, N-trimethylglycine) can be accumulated to high intracellular concentrations, conferring enhanced osmo- and cryotolerance upon Listeria monocytogenes. We report the identification of betL, a gene encoding a glycine betaine uptake system in L. monocytogenes, isolated by functional complementation of the betaine uptake mutant Escherichia coli MKH13. The betL gene is preceded by a consensus sigmaB-dependent promoter and is predicted to encode a 55 ...[more]

PMID: 10224004

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Project description:Salt stress is a common abiotic stress that limits the growth, development and yield of maize (Zea mays L.). To better understand the response of maize to salt stress and the mechanism by which exogenous glycine betaine (GB) alleviates the damaging effects of salt stress, the morphology, physiological and biochemical indexes, and root transcriptome expression profiles of seedlings of salt-sensitive inbred line P138 and salt-tolerant inbred line 8723 were compared under salt stress and GB-alleviated salt stress conditions. The results showed that under salt stress the growth of P138 was significantly inhibited and the vivo ion balance was disrupted, whereas 8723 could prevent salt injury by maintaining a high ratio of K+ to Na+. The addition of a suitable concentration of GB could effectively alleviate the damage caused by salt stress, and the mitigating effect on salt-sensitive inbred line P138 was more obvious than that on 8723. Transcriptome analysis revealed that 219 differentially expressed genes (DEGs) were up-regulated and 153 DEGs were down-regulated in both P138 and 8723 under NaCl treatment, and that 487 DEGs were up-regulated and 942 DEGs were down-regulated in both P138 and 8723 under salt plus exogenous GB treatment. In 8723 the response to salt stress is mainly achieved through stabilizing ion homeostasis, strong signal transduction activation, increasing reactive oxygen scavenging. GB alleviates salt stress in maize mainly by inducing gene expression changes to enhance the ion balance, secondary metabolic level, reactive oxygen scavenging mechanism, signal transduction activation. In addition, the transcription factors involved in the regulation of salt stress response and exogenous GB mitigation mainly belong to the MYB, MYB-related, AP2-EREBP, bHLH, and NAC families. We verified 10 selected up-regulated DEGs by quantitative real-time polymerase chain reaction (qRT-PCR), and the expression results were basically consistent with the transcriptome expression profiles. Our results from this study may provide the theoretical basis for determining maize salt tolerance mechanisms and the mechanism by which GB regulates salt tolerance.

Dataset Information

Identification and disruption of BetL, a secondary glycine betaine transport system linked to the salt tolerance of Listeria monocytogenes LO28.

Publications

Identification and disruption of BetL, a secondary glycine betaine transport system linked to the salt tolerance of Listeria monocytogenes LO28.

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