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Volume 155, Issue 1

Characterization of the LacI-type transcriptional repressor RbsR controlling ribose transport in ATCC 13032 Free

Abstract

The gene products of the () operon of (–) encode a ribose-specific ATP-binding cassette (ABC) transport system and its corresponding regulatory protein (RbsR). Deletion of the structural genes prohibited ribose uptake. Deletion of the regulatory gene resulted in an increased mRNA level of the whole operon. Analysis of the promoter region of the operon by electrophoretic mobility shift assays identified a catabolite-responsive element ()-like sequence as the RbsR-binding site. Additional RbsR-binding sites were identified in front of the recently characterized operon (---) and the ribokinase gene . , the repressor RbsR bound to its targets in the absence of an effector. A probable negative effector of RbsR is ribose 5-phosphate or a derivative thereof, since in a ribokinase ( ) double mutant, no derepression of the operon in the presence of ribose was observed. Analysis of the ribose stimulon in the wild-type revealed transcriptional induction of the and operons as well as of the gene. The inconsistency between the existence of functional RbsR-binding sites upstream of the ribokinase genes, their transcriptional induction during growth on ribose, and the missing induction in the mutant suggested the involvement of a second transcriptional regulator. Simultaneous deletion of the regulatory genes and finally demonstrated a transcriptional co-control of the and operons and the gene by both regulators, RbsR and UriR, which were furthermore shown to recognize the same cognate DNA sequences in the operators of their target genes.

  • Received:
  • Accepted:
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  • Published Online:
Keyword(s): ABC, ATP-binding cassette , CDS, coding sequence , EMSA, electrophoretic mobility shift assay , IMPACT, intein-mediated purification with an affinity chitin-binding tag and RACE, rapid amplification of cDNA ends
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2009-01-01
2024-04-23
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Characterization of the LacI-type transcriptional repressor RbsR controlling ribose transport in Corynebacterium glutamicum ATCC 13032
Microbiology 155, 150 (2009); https://doi.org/10.1099/mic.0.020388-0
/content/journal/micro/10.1099/mic.0.020388-0
/content/journal/micro/10.1099/mic.0.020388-0
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