Osmo-sensing by N- and C-terminal extensions of the glycine betaine uptake system BetP of Corynebacterium glutamicum.
The major uptake carrier for the compatible solute glycine betaine in Corynebacterium glutamicum is the secondary transport system BetP. It is effectively regulated by the external osmolality both on the level of expression and of activity. BetP carries highly charged domains both at the N and at the C terminus. We investigated the role of these extensions in the regulatory response to hyperosmotic stress. Mutants of the betP gene coding for proteins with truncated N- and C-terminal extensions were expressed in the C. glutamicum betP deletion strain DHP1 and were functionally characterized with respect to regulation of activity. The optimum of activation at 1.3 osmol/kg in wild type was shifted in the recombinant strains to about 2.6 osmol/kg in mutants with deletions in the N-terminal part. Deletions in the C-terminal domain resulted in a complete loss of regulation. The altered response to changes in osmolality led to severe consequences in the cellular adaption to hyperosmotic stress. Whereas in the wild type, the steady state level of glycine betaine accumulation is maintained by activity regulation of the BetP system itself, in the mutant with BetP proteins carrying truncations in the C-terminal domain, the observed steady state betaine accumulation was found to be due to a kinetic balance of unregulated glycine betaine uptake by the modifed BetP and efflux via the mechanosensitive efflux channel for compatible solutes at the same time.
Peter
H
H
Institut für Biotechnologie 1, Forschungszentrum Jülich GmbH, D-52425 Jülich, Germany.
Burkovski
A
A
Krämer
R
R
D000599:Amino Acids, Diamino / Q000494:pharmacology; D000779:Anesthetics, Local / Q000494:pharmacology; D001426:Bacterial Proteins; D001622:Betaine / Q000378:metabolism*; D001692:Biological Transport / Q000187:drug effects; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002414:Cations, Monovalent / Q000494:pharmacology; D003352:Corynebacterium / Q000187:drug effects / Q000502:physiology*; D015964:Gene Expression Regulation, Bacterial; D008958:Models, Molecular; D009154:Mutation; D009994:Osmolar Concentration; D009997:Osmotic Pressure; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000378:metabolism; D015398:Signal Transduction; D012964:Sodium / Q000494:pharmacology; D012965:Sodium Chloride / Q000494:pharmacology; D013012:Sorbitol / Q000494:pharmacology; D027981:Symporters; D013748:Tetracaine / Q000494:pharmacology
D000599:Amino Acids, Diamino; D000779:Anesthetics, Local; D001426:Bacterial Proteins; C101652:BetP protein, Corynebacterium glutamicum; D002352:Carrier Proteins; D002414:Cations, Monovalent; D010446:Peptide Fragments; D011994:Recombinant Proteins; D027981:Symporters; D013748:Tetracaine; D001622:Betaine; D012965:Sodium Chloride; D013012:Sorbitol; C045628:ectoine; D012964:Sodium