1887

Abstract

The operon encodes three iron–sulfur-containing proteins required for -lactate utilization and involved in biofilm formation. The transcriptional regulator LutR of the GntR family negatively controls expression. The gene, which is situated immediately upstream of , encodes an -lactate permease. Here, we show that expression can be strongly induced by -lactate and is subject to partial catabolite repression by glucose. Disruption of the gene led to a strong derepression of and no further induction by -lactate, suggesting that the LutR repressor can also negatively control expression. Electrophoretic mobility shift assay revealed a LutR-binding site located downstream of the promoter of or and containing a consensus inverted repeat sequence 5′-TCATC-N-GATGA-3′. Reporter gene analysis showed that deletion of each LutR-binding site caused a strong derepression of or . These results indicated that these two LutR-binding sites can function as operators . Moreover, deletion analysis identified a DNA segment upstream of the promoter to be important for expression. In contrast to the truncated LutR of laboratory strains 168 and PY79, the full-length LutR of the undomesticated strain RO-NN-1, and probably many other strains, can directly and negatively regulate transcription. The absence or presence of the N-terminal 21 aa of the full-length LutR, which encompass a small part of the predicted winged helix–turn–helix DNA-binding motif, may probably alter the DNA-binding specificity or affinity of LutR.

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2014-10-01
2020-09-19
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