Proline Utilization by Bacillus subtilis : Uptake and Catabolism Susanne Moses a , Tatjana Sinner a , Adrienne Zaprasis a , Nadine Stöveken a , Tamara Hoffmann a , Boris R. Belitsky b , Abraham L. Sonenshein b and Erhard Bremer a a Philipps-University Marburg, Department of Biology, Laboratory for Microbiology, Marburg, Germany b Tufts University School of Medicine, Department of Molecular Biology and Microbiology, Boston, Massachusetts, USA ABSTRACT l -Proline can be used by Bacillus subtilis as a sole source of carbon or nitrogen. We traced l -proline utilization genetically to the putBCP ( ycgMNO ) locus. The putBCP gene cluster encodes a high-affinity proline transporter (PutP) and two enzymes, the proline dehydrogenase PutB and the Δ 1 -pyrroline-5-carboxylate dehydrogenase PutC, which jointly catabolize l -proline to l -glutamate. Northern blotting, primer extension, and putB-treA reporter gene fusion analysis showed that the putBCP locus is transcribed as an l -proline-inducible operon. Its expression was mediated by a SigA-type promoter and was dependent on the proline-responsive PutR activator protein. Induction of putBCP expression was triggered by the presence of submillimolar concentrations of l -proline in the growth medium. However, the very large quantities of l -proline (up to several hundred millimolar) synthesized by B. subtilis as a stress protectant against high osmolarity did not induce putBCP transcription. Induction of putBCP transcription by external l -proline was not dependent on l -proline uptake via the substrate-inducible PutP or the osmotically inducible OpuE transporter. It was also not dependent on the chemoreceptor protein McpC required for chemotaxis toward l -proline. Our findings imply that B. subtilis can distinguish externally supplied l -proline from internal l -proline pools generated through de novo synthesis. The molecular basis of this regulatory phenomenon is not understood. However, it provides the B. subtilis cell with a means to avoid a futile cycle of de novo l -proline synthesis and consumption by not triggering the expression of the putBCP l -proline catabolic genes in response to the osmoadaptive production of the compatible solute l -proline.
Proline Utilization by Bacillus subtilis: Uptake and Catabolism
Abstract
Proline Utilization by Bacillus subtilis : Uptake and Catabolism Susanne Moses a , Tatjana Sinner a , Adrienne Zaprasis a , Nadine Stöveken a , Tamara Hoffmann a , Boris R. Belitsky b , Abraham L. Sonenshein b and Erhard Bremer a a Philipps-University Marburg, Department of Biology, Laboratory for Microbiology, Marburg, Germany b Tufts University School of Medicine, Department of Molecular Biology and Microbiology, Boston, Massachusetts, USA ABSTRACT l -Proline can be used by Bacillus subtilis as a sole source of carbon or nitrogen. We traced l -proline utilization genetically to the putBCP ( ycgMNO ) locus. The putBCP gene cluster encodes a high-affinity proline transporter (PutP) and two enzymes, the proline dehydrogenase PutB and the Δ 1 -pyrroline-5-carboxylate dehydrogenase PutC, which jointly catabolize l -proline to l -glutamate. Northern blotting, primer extension, and putB-treA reporter gene fusion analysis showed that the putBCP locus is transcribed as an l -proline-inducible operon. Its expression was mediated by a SigA-type promoter and was dependent on the proline-responsive PutR activator protein. Induction of putBCP expression was triggered by the presence of submillimolar concentrations of l -proline in the growth medium. However, the very large quantities of l -proline (up to several hundred millimolar) synthesized by B. subtilis as a stress protectant against high osmolarity did not induce putBCP transcription. Induction of putBCP transcription by external l -proline was not dependent on l -proline uptake via the substrate-inducible PutP or the osmotically inducible OpuE transporter. It was also not dependent on the chemoreceptor protein McpC required for chemotaxis toward l -proline. Our findings imply that B. subtilis can distinguish externally supplied l -proline from internal l -proline pools generated through de novo synthesis. The molecular basis of this regulatory phenomenon is not understood. However, it provides the B. subtilis cell with a means to avoid a futile cycle of de novo l -proline synthesis and consumption by not triggering the expression of the putBCP l -proline catabolic genes in response to the osmoadaptive production of the compatible solute l -proline.Preview Only. This article cannot be rented because we do not currently have permission from the publisher.
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Proline Utilization by Bacillus subtilis: Uptake and Catabolism
Moses, Susanne; Sinner, Tatjana; Zaprasis, Adrienne; Stöveken, Nadine; Hoffmann, Tamara; Belitsky, Boris R.; Sonenshein, Abraham L.; Bremer, Erhard
Follow Journal of Bacteriology
, Volume 194 (4): 745
American Society For Microbiology – Feb 15, 2012
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