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PMID:21964733
| Citation |
Huang, SC, Lin, TH and Shaw, GC (2011) PrcR, a PucR-type transcriptional activator, is essential for proline utilization and mediates proline-responsive expression of the proline utilization operon putBCP in Bacillus subtilis. Microbiology (Reading, Engl.) 157:3370-7 |
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| Abstract |
The soil bacterium Bacillus subtilis can utilize exogenous proline as a sole nitrogen or carbon source. The proline-inducible putBCP (formerly ycgMNO) operon encodes proteins responsible for proline uptake and two-step oxidation of proline to glutamate. We now report that a gene (formerly ycgP, now designated prcR) located downstream of the putBCP operon is essential for B. subtilis cells to utilize proline as a sole nitrogen or carbon source. Disruption of the prcR gene also abolished proline induction of putB transcription. prcR expression is not subject to autoregulation and proline induction. The PrcR protein shows no significant amino acid sequence similarity to the known transcriptional activators for proline utilization genes of other bacteria, but it does show partial amino acid sequence similarity to the transcriptional regulator PucR for the purine degradation genes of B. subtilis. PrcR orthologues of unknown function are present in some other Bacillus species. Primer-extension analysis suggests that both putB and prcR are transcribed by a σ(A)-dependent promoter. Deletion and mutation analysis revealed that an inverted repeat (5'-TTGTGG-N5-CCACAA-3') centred at position -76 relative to the transcriptional initiation site of putB is essential for putB expression. Electrophoretic mobility shift assays showed that the purified His-tagged PrcR was capable of binding specifically to this inverted repeat. Altogether, these results suggest that PrcR is a PucR-type transcriptional activator that mediates expression of the B. subtilis putBCP operon in response to proline availability. |
| Links |
PubMed Online version:10.1099/mic.0.054197-0 |
| Keywords |
Bacillus subtilis/genetics; Bacillus subtilis/growth & development; Bacillus subtilis/metabolism; Binding Sites; Carbon/metabolism; DNA Mutational Analysis; Electrophoretic Mobility Shift Assay; Gene Deletion; Gene Expression Regulation, Bacterial; Membrane Transport Proteins/biosynthesis; Nitrogen/metabolism; Operon; Proline/metabolism; Promoter Regions, Genetic; Protein Binding; Sequence Homology, Amino Acid; Sigma Factor/metabolism; Trans-Activators/genetics; Trans-Activators/metabolism; Transcription, Genetic |
| edit table |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| GO:0051699: proline oxidase activity |
IMP: Inferred from Mutant Phenotype: |
F |
Figure 2 C. This figure shows how the prcR mutant stops the expression of putB. Without the transcriptional activator, prcR, putB cannot be transcribed and L-proline cannot be utilized by the cell as a nutrient source. |
complete |
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| GO:0051699: proline oxidase activity |
IMP: Inferred from Mutant Phenotype: |
F |
Figure 2b: Results from the transcriptional analysis of putB expression by RT-PCR with putB-specific primers and 16S rRNA-specific primers. putB is expressed in the wild type, but not in the prcR mutant. Without the transcriptional activator, prcR, putB is not transcribed. This means that L-proline cannot be utilized as a nutrient source. |
complete |
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See also
References
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