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PMID:10347192
| Citation |
Bird, TH, Du, S and Bauer, CE (1999) Autophosphorylation, phosphotransfer, and DNA-binding properties of the RegB/RegA two-component regulatory system in Rhodobacter capsulatus. J. Biol. Chem. 274:16343-8 |
|---|---|
| Abstract |
In the purple, photosynthetic bacterium, Rhodobacter capsulatus, the RegB/RegA two-component system is required for activation of several anaerobic processes, such as synthesis of the photosynthetic apparatus and assimilation of CO2 and N2. It is believed that RegB is an integral membrane histidine kinase that monitors the external environment. Under anaerobic growth conditions, it transduces a signal through phosphorylation of the response regulator, RegA, which then induces target gene expression. We used an in vitro assay to characterize the phosphorylation of wild-type RegA and a mutant variant (RegA*) that is responsible for abnormally high photosynthesis gene expression under both aerobic and anaerobic growth conditions. Phosphorylation assays indicate that phosphorylated RegA* (RegA* approximately P) is much more stable than RegA approximately P, indicating that it may be locked in a conformation that is resistant to dephosphorylation. DNase I footprint assays also indicate that unphosphorylated RegA* has a much higher affinity for specific DNA binding sites than the wild-type protein. Phosphorylation of RegA* increases DNA binding 2. 5-fold, whereas phosphorylation of RegA increases DNA binding more than 16-fold. Collectively, these results support the hypothesis that RegA* is a constitutively active variant that does not require phosphorylation to assume a structural conformation required to bind DNA. |
| Links | |
| Keywords |
Bacterial Proteins/metabolism; DNA, Bacterial/metabolism; Deoxyribonuclease I/metabolism; Phosphorylation; Photosynthetic Reaction Center Complex Proteins/metabolism; Promoter Regions, Genetic; Protein Conformation; Protein Kinases; Rhodobacter capsulatus/metabolism; Structure-Activity Relationship; Transcription Factors/metabolism |
| edit table |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
|
enables |
GO:0000156: phosphorelay response regulator activity |
ECO:0000314: direct assay evidence used in manual assertion |
F |
Seeded From UniProt |
complete | |||
|
involved_in |
GO:0000160: phosphorelay signal transduction system |
ECO:0000314: direct assay evidence used in manual assertion |
P |
Seeded From UniProt |
complete | |||
| GO:0000160 : two-component signal transduction system (phosphorelay) |
ECO:0000314: |
P |
Figures 2, 3 |
complete | ||||
| GO:0000156: two-component response regulator activity |
ECO:0000314: |
F |
Figure 5 shows DNA affinity |
complete | ||||
| GO:0016301: kinase activity |
ECO:0000314: |
F |
Figures 2,3 |
complete | ||||
| GO:0000160: two-component signal transduction system (phosphorelay) |
ECO:0000314: |
P |
Figures 2,3 |
complete | ||||
| GO:0046777: protein autophosphorylation |
ECO:0000314: |
P |
Figure 1 |
complete | ||||
|
enables |
GO:0016301: kinase activity |
ECO:0000314: direct assay evidence used in manual assertion |
F |
Seeded From UniProt |
complete | |||
|
involved_in |
GO:0000160: phosphorelay signal transduction system |
ECO:0000314: direct assay evidence used in manual assertion |
P |
Seeded From UniProt |
complete | |||
|
involved_in |
GO:0046777: protein autophosphorylation |
ECO:0000314: direct assay evidence used in manual assertion |
P |
Seeded From UniProt |
complete | |||
See also
References
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