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Volume 160, Issue 10

Functional characterization of the locus for -Glu and -Gln utilization in PAO1 Free

Abstract

-Glu, an essential component of peptidoglycans, can be utilized as a carbon and nitrogen source by . DNA microarrays were employed to identify genes involved in -Glu catabolism. Through gene knockout and growth phenotype analysis, the divergent – (-Glu utilization) gene cluster was shown to participate in -Glu and -Gln catabolism and regulation. Growth of the and mutants was abolished completely on -Glu or retarded on -Gln as the sole source of carbon and/or nitrogen. The gene encoded a FAD-dependent -amino acid dehydrogenase with -Glu as its preferred substrate, and its promoter was specifically induced by exogenous -Glu and -Gln. The function of DguR as a transcriptional activator of the operon was demonstrated by promoter activity measurements and by mobility shift assays with purified His-tagged DguR . Although the DNA-binding activity of DguR did not require -Glu, the presence of -Glu, but not -Gln, in the binding reaction was found to stabilize a preferred nucleoprotein complex. The presence of a putative DguR operator was revealed by analysis of the – intergenic regions among spp. and binding of DguR to a highly conserved 19 bp sequence motif was further demonstrated. The gene encodes a putative enamine/imine deaminase of the RidA family, but its role in -Glu catabolism remains to be determined. Whilst a lesion in encoding a periplasmic solute binding protein only affected growth on -Glu slightly, expression of the previously characterized AatJMQP transporter for acidic -amino acid uptake was found inducible by -Glu and essential for -Glu utilization. In summary, the findings of this study supported DguA as a new member of the FAD-dependent -amino acid dehydrogenase family, and DguR as a -Glu sensor and transcriptional activator of the promoter.

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  • Accepted:
  • Published Online:
Funding
This study was supported by the:
  • National Science Foundation (Award NSF0950217)
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2014-10-01
2025-01-26
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/content/journal/micro/10.1099/mic.0.081141-0
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Functional characterization of the dguRABC locus for d-Glu and d-Gln utilization in Pseudomonas aeruginosa PAO1
Microbiology 160, 2331 (2014); https://doi.org/10.1099/mic.0.081141-0
/content/journal/micro/10.1099/mic.0.081141-0
/content/journal/micro/10.1099/mic.0.081141-0
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