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PMID:12597891
| Citation |
Monteiro, RA, Souza, EM, Geoffrey Yates, M, Steffens, MB, Pedrosa, FO and Chubatsu, LS (2003) Expression, purification, and functional analysis of the C-terminal domain of Herbaspirillum seropedicae NifA protein. Protein Expr. Purif. 27:313-8 |
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| Abstract |
The Herbaspirillum seropedicae NifA protein is responsible for nif gene expression. The C-terminal domain of the H. seropedicae NifA protein, fused to a His-Tag sequence (His-Tag-C-terminal), was over-expressed and purified by metal-affinity chromatography to yield a highly purified and active protein. Band-shift assays showed that the NifA His-Tag-C-terminal bound specifically to the H. seropedicae nifB promoter region in vitro. In vivo analysis showed that this protein inhibited the Central + C-terminal domains of NifA protein from activating the nifH promoter of K. pneumoniae in Escherichia coli, indicating that the protein must be bound to the NifA-binding site (UAS site) at the nifH promoter region to activate transcription. |
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| Keywords |
Bacterial Proteins/chemistry; Bacterial Proteins/isolation & purification; Bacterial Proteins/physiology; Betaproteobacteria/metabolism; Chromatography; Cloning, Molecular; DNA/metabolism; Escherichia coli/metabolism; Klebsiella pneumoniae/genetics; Plasmids/metabolism; Promoter Regions, Genetic; Protein Binding; Protein Structure, Tertiary; Recombinant Fusion Proteins/metabolism; Transcription Factors/chemistry; Transcription Factors/isolation & purification; Transcription Factors/physiology; Transcription, Genetic; Transcriptional Activation |
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Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
|
involved_in |
GO:0045893: positive regulation of transcription, DNA-templated |
ECO:0000315: mutant phenotype evidence used in manual assertion |
P |
Seeded From UniProt |
complete | |||
| GO:0045893: positive regulation of transcription, DNA-templated |
ECO:0000315: |
P |
Table 1 |
complete | ||||
See also
References
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