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 DctD-mediated transcriptional activation from the R. leguminosarum dctA promoter both in vivo and in vitro. In contrast to R. leguminosarum dctA, the Sinorhizobium meliloti dctA promoter region was found to have a much weaker match 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 dtA that reduced the affinity of the promoter regulatory region for IHF by approximately 30-fold and resulted in an eight-fold decrease in transcriptional activation in both R. leguminosarum and S. meliloti. These data suggest that both rhizobial species have an IHF homolog that stimulates DctD-mediated transcriptional activation from the R. leguminosarum dctA promoter. Consistent with this hypothesis, a 12.5 kDa protein was identified from R. leguminosarum as a putative homolog of IHF subunit beta by immunoblotting and N-terminal sequence analysis.
Sojda
J
J
Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park 16802, USA.
Gu
B
B
Lee
J
J
Hoover
T R
TR
Nixon
B T
BT
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D029382:Dicarboxylic Acid Transporters; D034982:Integration Host Factors; D008969:Molecular Sequence Data; D010641:Phenotype; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D016961:Rhizobium leguminosarum / Q000235:genetics*; D016962:Sinorhizobium meliloti / Q000235:genetics*; D013045:Species Specificity; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D002352:Carrier Proteins; D017931:DNA Primers; D004268:DNA-Binding Proteins; D029382:Dicarboxylic Acid Transporters; D034982:Integration Host Factors; C073786:dctA protein, Sinorhizobium meliloti