A rhizobial homolog of IHF stimulates transcription of dctA in Rhizobium leguminosarum but not in Sinorhizobium meliloti

Sequence inspection identified several potential IHF binding sites adjacent to the Rhizobium leguminosarum dctA promoter. IHF protected the -30 to -76 region from DNase I digestion, but systematic error in quantitative assays suggested that this protein-DNA interaction is complex. IHF stimulated Dct D-mediated transcriptional activation from the R. leguminosarum dctA promot er both in vivo and in vitro. In contrast to R. leguminosarum dctA, the Sin orhizobium meliloti dctA promoter region was found to have a much weaker ma tch to the consensus IHF binding site and a low affinity for IHF. Moreover, IHF had no effect on transcriptional activation from the S. meliloti dctA promoter in vitro. A base substitution was introduced into the IHF binding site of R. leguminosarum dctA that reduced the affinity of the promoter reg ulatory region for IHF by similar to 30-fold and resulted in an eight-fold decrease in transcriptional activation in both R. leguminosarum and S. meli loti. These data suggest that both rhizobial species have an IHF homolog th at stimulates DctD-mediated transcriptional activation from the Ii. legumin osarum dctA promoter. Consistent with this hypothesis, a 12.5 kDa protein w as identified from R. leguminosarum as a putative homolog of IHF subunit be ta by immunoblotting and N-terminal sequence analysis. (C) 1999 Elsevier Sc ience B.V. All rights reserved.