Ntr-like promoters and upstream regulatory sequence ftr are required for transcription of a developmentally regulated Caulobacter crescentus flagellar gene.
The flbG (hook operon or transcription unit II) and flaN (transcription unit I) operons of Caulobacter crescentus have a -12, -24 nucleotide sequence motif that is very similar to those of the Nif and Ntr promoters of enteric bacteria and Rhizobium spp. and a conserved ftr (flagellar gene transcription regulation) sequence, previously designated II-1 (D. A. Mullin, S. A. Minnich, L.-S. Chen, and A. Newton, J. Mol. Biol. 195:939-943, 1987) at approximately -100. We have used site-directed mutagenesis to examine the role of these sequences in the transcriptional regulation of these periodically expressed flagellar genes. Mutations in the flbG promoter that removed the conserved GC at -12, -13, the GG at -24, -25, or an AC base pair at -18, -19 in the nonconserved sequence between the -12, -24 elements completely eliminated detectable transcription. Mutations at other positions resulted in either a slight decrease (position 26), no change (position 15), or an elevated level (position -16 or -19) of the flbG transcript. By contrast, most of these flbG promoter mutations resulted in greatly elevated levels of transcription from the opposing flaN operon. Similar experiments were used to confirm the location of the flaN promoter to a -12, -24 Nif and Ntr sequence motif. Deletion of all or part of the ftr element or point mutations in the sequence drastically reduced the level of flbG transcript and resulted in increased levels of the flaN transcript. Thus, the conserved sequences at -12 and -24 in flbG and flaN are required for transcription of these genes in vivo, and the ftr element is required for transcription of flbG. This analysis also suggested that the ftr sequence and sequences in the flbG promoter are required for the negative autoregulation of the flbG and flaN operons. We speculate that the flbG and flaN promoters and the ftr element interact in some way to mediate the negative control of these divergent transcription units.
Mullin
D A
DA
Department of Biology, Tulane University, New Orleans, Louisiana 70118.
Newton
A
A
D001483:Base Sequence; D004252:DNA Mutational Analysis; D004269:DNA, Bacterial / Q000235:genetics; D005407:Flagella / Q000502:physiology*; D005408:Flagellin / Q000235:genetics; D005786:Gene Expression Regulation; D005798:Genes, Bacterial; D006090:Gram-Negative Bacteria / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012329:RNA, Bacterial / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping
D004269:DNA, Bacterial; D012329:RNA, Bacterial; D005408:Flagellin